Heme oxygenase-1 alleviates ischemia/reperfusion injury in aged liver

AIM: To investigate if ischemia/reperfusion (I/R) injury in aged liver could be alleviated by heme oxygenase-1 (HO-1).METHODS: Three groups of SD rats (16 mo old) were studied. Group 1: control donors received physiological saline 24 h before their livers were harvested; group 2: donors were pretreated with hemih 24 h before their livers were harvested; and group 3: donors received hemin 24 h before their livers were harvested and zinc protoporphyrin (ZnPP,HO-1 inhibitor) was given to recipients at reperfusion. The harvested livers were stored in University of Wisconsin solution (4 ℃) for 6 h, and then transplanted to syngeneic rats. Serum glutamic oxaloacetic transaminase (SGOT),apoptotic cells, and apoptotic gene were measured 3, 6,12, 24, 48 h after reperfusion. We measured the apoptotic index by TUNEL, determined the expression of antiapoptotic Bcl-2 and proapoptotic (caspase-3) gene products by Western blot.RESULTS: After 3, 6, 12, 24, and 48 h of reperfusion, the SGOT levels (584.4±85.8 u/L, 999.2±125.2 u/L, 423.4±161.3u/L, 257.8±95.8 u/L, and 122.4±26.4 u/L) in hemin group were significantly (all P＜0.05) lower than those in saline group (1082.2±101.2 u/L, 1775.2±328.3 u/L, 840.4±137.8 u/L,448.6±74.3 u/L, and 306.2±49.3 u/L). Liver HO-1 enzymatic activity correlated with beneficial effects of hemin and deleterious effects of adjunctive ZnPP treatment. Markedly less apoptotic (TUNEL+) liver cells 3, 6, 12, 24, and 48 h after reperfusion (5.16±0.73, 10.2±0.67, 9.28±0.78, 7.14±1.12,and 4.78±0.65) (P＜0.05) could be detected in hemin liver grafts, as compared to controls (7.82±1.05, 15.94±1.82,11.67±1.59, 8.28±1.09, and 6.36±0.67). We detected the increased levels of Bcl-2 (1.5-fold) expression and compared with saline controls. These differences were most pronounced at 12 h after transplantation. In contrast, an active form of proapoptotic caspase-3 (p20) protein was found to be 2.9-fold lower at 24 h in hemin-pretreated group, as compared to saline liver transplant controls.CONCLUSION: HO-1 overexpression can provide potent protection against cold I/R injury. This effect depends, at least in part, on HO-1-mediated inhibition of antiapoptotic mechanism.

Gastroprotective effect and mechanism of amtolmetin guacyl in mice

AIM: To investigate the gastroprotective effect and mechanism of amtolmetin guacyl (AMG, MED15) in mice.METHODS: Male and female Kunming strain mice,weighing 18-22 g, were utilized in the experiment. Normal or ethanol-induced gastric mucosal damage models in mice were successfully established to investigate the gastroprotective effect and mechanism of AMG. In the experiment of gastric mucosal damage after repeated treatment with AMG, the mice were randomly divided into 5 groups: normal group, 3 AMG groups receiving (75, 150 and 300 mg/kg), and tolmetin group receiving 90 mg/kg.The mice were randomly divided into 6 groups as follows:normal group, model group, AMG groups with doses of 75, 150 and 300 mg/kg, respectively, and tolmetin group with a dose of 90 mg/kg in ethanol-induced gastric mucosal damage experiment. The severity of gastric mucosal lesions was scored from 0 to 5. Gastric tissue sections were stained with hematoxylin and eosin (HE) and examined under light microscopy. Also gastric tissue sections were stained with uranyl acetate and lead citrate, and examined under electron microscopy. In addition, nitric oxide (NO) and malondialdehyde (MDA) contents, and nitric oxide synthase (NOS) and superoxide dismutase (SOD) activities in the stomach tissue homogenates were measured by biochemical methods.RESULTS: Repeated treatment with AMG (75, 150 and3 00 mg/kg) for 7 d did not induce any appreciable mucosal damage, and the average score was not significantly different from that of normal mice. In contrast, tolmetin (90 mg/kg) produced significant gastric mucosal lesions compared with the normal group (P<0.01). AMG (75, 150 and 300 mg/kg) significantly reduced the severity of gastric lesions induced by ethanol in a dose-dependent manner as compared with the model group (P<0.05, AMG 75 and 150 mg/kg vs model; P<0.01, AMG 300 mg/kg vs model).Light and electron microscopy revealed that AMG (150 and 300 mg/kg) induced minimal changes in the surface epithelium layer, without vascular congestion or leucocyte adherence. AMG (75,150 and 300 mg/kg) demonstrated dose-dependent gastroprotective effects on mice in ourstudy. AMG (75, 150 and 300 mg/kg) could significantly increase NO content and NOS level in the stomach homogenates of mice compared with the model group (P<0.05, AMG 75 mg/kg and 150 mg/kg groups vs model group; P<0.01, AMG 300 mg/kg vs model group) respectively. Moreover, AMG (150 and 300 mg/kg) not only significantly increased SOD activities but also obviously decreased the MDA content in the stomach homogenates of mice.CONCLUSION: AMG exerts significant gastroprotective actions on mice and the involved mechanisms may be its antioxidative effect and induction of NO production.

Expression of intestinal trefoil factor, proliferating cell nuclear antigen and histological changes in intestine of rats after intrauterine asphyxia

AIM: To study the expressions of intestinal trefoil factor (ITF) and proliferating cell nuclear antigen (PCNA) and histologic changes in intestine, to investigate the relationship between ITF and intestinal damage and repair after intrauterine hypoxia so as to understand the mechanism of intestinal injury and to find a new way to prevent and treat gastrointestinal diseases.METHODS: Wistar rats, pregnant for 21 d, were used to establish animal models of intrauterine asphyxia by clamping one side of vessels supplying blood to uterus for 20 min, another side was regarded as sham operation group. Intestinal tissues were taken away at 0, 24, 48and 72 h after birth and stored in different styles. ITF mRNA was detected by RT-PCR. PCNA expression was measured by immunohistochemistry. Intestinal tissues were studied histologically by HE staining in order to observe the areas and degree of injury and to value the intestinal mucosa injury index (IMDI).RESULTS: ITF mRNA appeared in full-term rats and increased with age. After ischemia, ITF mRNA was decreased to the minimum (0.59±0.032) 24 h after birth, then began to increase higher after 72 h than it was in the control group (P＜0.01). PCNA positive staining located in goblet cell nuclei. The PCNA level had a remarkable decline (53.29±1.97) 48 h after ischemia. Structure changes were obvious in 48-h group, IMDI (3.40±0.16) was significantly increased. Correlation analyses showed that IMDI had a negative correlation with ITF mRNA and PCNA (r = -0.543,P＜0.05; r = -0.794, P＜0.01, respectively).CONCLUSION: Intrauterine ischemia can result in an early decrease of ITF mRNA expression. ITF and PCNA may play an important role in the damage and repair of intestinal mucosa.

Association of HLA-B Alleles With Human Immunodeficiency Virus Type 1 Infection in the Yi Ethnic Group in Sichuan Province

Objective To determine the distribution of HLA-B alleles in the Chinese Yi ethnic group and its association with HIV infection. Methods One hundred and six unrelated healthy HIV negative and 73 HIV positive Chinese Yi ethnic individuals were typed by PCR-SSP. Results The frequency of alleles B*07, B*35, and B*46 were increased in HTV-1 -positive subjects, whereas the alleles B*55, B*44 and B*78 were absent in the HIV-infected persons studied. The B*46 allele was present in a significantly higher gene frequency among HIV-1-positive individuals (P=0.02, OR=3.32, 95% CI=1.13-9.78) compared with control subjects. Conclusion HLA-B*46 may be associated with its susceptibility to HIV-1 infections.

Dynamic change of epidermal growth factor in neonatal rat with intestine injury

AIM: To determine whether diminished levels of epidermal growth factor (EGF) were present in neo-natal rats with intestinal injury and related with the degree of intestinal injury, so we modeled a model in neo-natal rats of intestinal injury and to examine the dynamic levels of EGF on injury of intestine.METHODS: One-day-old Wistar rat pups received an intraperitoneally injection with 4 mg/kg lipopolysaccharide (LPS), followed by collection of ileum tissue at 1, 3, 6, 12,and 24 h following LPS administration. The ileum was for histological evaluation of NEC and for measurements of EGF using ABC-ELISA. The correlation between the degree of intestinal injury and levels of EGF was determined. RESULTS: The LPS-injected pups also showed a significant increase in injury scores at 1, 3, 6, 12, and 24 h [respectively, (1.08±0.61), (1.63±0.84), (1.95±0.72), (2.42±0.43)and (2.21±0.53)] vsthe control (0.12±0.17) (P＜0.01).EGF levels at 1, 3, 6, 12 h [respectively, (245.6±49.0), (221.4±39.0), (223.4±48.1), (246.0±46.6)] pg/mg were significantly loss than the control (275.6±50.4) pg/mg (P＜0.05). There was a significant negative correlation between the EGF levels and the grade of intestinal injury within 24 h (P＜0.05).CONCLUSION: Neo-natal rats with intestinal injury have significantly lower levels of ileum EGF. Reduced levels of this growth factor might be related to the pathogenesis of NEC.

FR167653 attenuates murine immunological liver injury

AIM: To study the effect of FR167653 on immunologica liver injury (ILI) in mice.METHODS: ILI was established by tail vein injection of 2.5 mg Bacillus Calmette-Guerin(BCG), and 10 d later with 10 mg lipopolysaccharide (LPS) in 0.2 mL saline (BCG plus LPS). Alanine aminotransferase (ALT), aspartate aminotransferase (AST) in sera and malondialdehyde (MDA), glutathione peroxidase (GSHpx) contents in liver homogenates were assayed by spectrophotometry. The levels of tumor necrosis factor-~ (TNF-~) and nitric oxide (NO) levels in sera were determined using ELISA. Interleukin-1 (IL-1) produced by peritoneal macrophages was determined by the method of 3^H-infiltrated cell proliferation. The nuclear factor-kappa B (NF-xB) p65 in liver tissue was analyzed with reverse transcription polymerase chain reaction (RT-PCR). Liver samples collected were stained with hematoxylin and eosin.RFSULTS: FR167653 (50, 100, 150 mg/kg) could significantly decrease the serum transaminase (ALl', AS'I-) activity and MDA content in liver homogenate, and improve reduced GSHpx level of liver homogenate. Liver histopathological examinatdon showed FR167653 (100, 150 mg/kg) significantly reduced inflammatory cells infiltration and liver cells necrosis. FR167653 (50, 100, 150 mg/kg) significantly lowered TNF-α and NO levels in serum, and IL-1 produced by peritoneal macrophages. Moreover, expression of NF-xBmRNA in liver tissue of ILl induced by BCG plus LPS was significantly reduced by FR167653.CONCLUSION: All results showed that FR167653 had significant inhibitorv action an ILI in mice.

Separation of Aniline Derivatives by Micellar Electrokinetic Capillary Chromatography

A micellar electrokinetic capillary chromatography (MECC) was developed for the determination of aniline and 6 substituted anilines. The seven components were separated within 25 min in the buffer solution of 40 mmol/L sodium borate and 100 mmol/L SDS. It was found that the separation was dependent on operating voltage, pH value, borate and SDS concentrations. The analytical performance was examined in terms of linear response and reproducibility. Wastewater was determined by the established method.

Prognosis-Related Analysis of Renal Pelvic Cancer

OBJECTIVE To promote the diagnosis and therapeutic results for renalpelvic cancer. METHODS The prognosis-related factors in 47 cases with renal pelvic cancer were analyzed retrospectively. RESULTS The overall 3 and 5-year survival rates for renal pelvic cancer patients were 65.9% (31/47) and 51.1% (24/47), respectively. The 5-year survival was 55% (23/40) in organ-confined cancer and 26.7% (2/7) with coexisting multi-organ involvement (P>0.05). The 5-year survival was 38.7%(12/31) in cases with a tumor >2.5 cm and 75%(12/16) in the cases with tumor ≤2.5 cm (P<0.05). The 5-year survival was 37.9% (11/29) in cases with serious hydronephrosis, which was significantly lower than the 72.2% (13/18) found in those with slight hydronephrosis (P <0.05). According to the histological grade, the 5-year survival was 100% (6/6) in patients with a G1 tumor, 65.2% (15/23) with G2, and 16.7% (3/18) with G3 (P<0.01). Based on the pathologic stage, the 5-year survival of cases was 84.6% (11/13)with T1 tumors, 60% (12/20)with T2, and 7.1% (1/14) with T3～T4 (P<0.01). Patients with a G2T2 or higher staging tumor, Who underwent radical nephroureterectomy with partial bladder resection by a transabdominal approach had a significantly higher 5-year survival than those who underwent nephrectomy or nephroureterectomy with partial bladder resection via a lumbar approach (P <0.05). There was no significant difference between the 5-year survival of patients with recurrence of bladder carcinoma compared to patients without recurrence (P>0.05).CONCLUSION The tumor grade and stage are the key points for prognosis. Radical nephroureterectomy with partial bladder resection is an effective method to improve the prognosis of patients with a high grade and high stage tumor.

Leflunomide attenuates hepatocyte injury by inhibiting Kupffer cells

AIM: To investigate the importance of direct contact between Kupffer cells (KCs) and hepatocytes (HCs) during hepatic inflammatory responses, and the effect of leflunomide's active metabolite, A771726, on cytokines in KCs, HCs and KC cocultures (DC cocultures). METHODS: KCs and HCs in liver were isolated by digestion with pronase and collagenase. Lipopolysaccharide (LPS)-induced inflammatory response in monocultures of rat HCs and KCs was compared with that in DC cocultures. Tumorn ecrosis factor-α (TNF-α） and interleukin-1 (IL-1) concentrations in different culture supernatants were measured with ELISA. TNF-α mRNA in KCs of inflammatory liver injury was analyzed with reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: DC cocultures strongly exhibited the production of TNF-α and IL-1 compared with other cultures, and these cytokines were mainly produced by KCs, especially byactivated KCs. Time course studies revealed an increased production of TNF-α preceding the IL-1 production, suggesting that increased TNF-α levels could be involved in the increase of IL-1 production. Leflunomide's active metabolite, A771726, had significantly inhibitory effect on TNF-α and IL-1 at protein and transcription levels, and the reduced production of IL-1 by A771726 was associated with the inhibitory action of A771726 on TNF-α. CONCLUSION: Leflunomide can inhibit hepatoolte damage by inhibiting proinflammatory cytokine release from KCs.

The Role of Innate Immune Cells in the Response of Heat-Treated Mycobacterium tuberculosis (M.tb) Antigens Stimulating PBMCs

The proliferation response of γδT cells to the antigen from heat-treated Mycobacterium tuberculosis H37Ra (M.tb Ag)was used as a good model in γδT cell research.From preliminary research it is found that activated NK cells positively elevated γδT cells proliferation after simulating PBMCs with M.tb Ag.To investigate different behaviors of NK cells,γδNKT cells,γδT cells and relationships between these cell subsets,activation and proliferation of different cell subsets of PBMCs in response to M.tb Ag were analyzed.We demonstrated that NK cells,γδNKT cells and γδT cells could be activated after stimulation with M.tb Ag.γδNKT cells and γδT cells proliferated while the number of NK cells decreased after 11 day-simulation with M.tb Ag.Meanwhile,at the early time of stimulation the cytotoxicity of PBMCs was enhanced.Cellular & Molecular Immunology. 2004;1(6):467-470.

Gene Identification and Expression Analysis of 86,136 Expressed Sequence Tags(EST)from the Rice Genome

Expressed Sequence Tag (EST) analysis has pioneered genome-wide gene discovery and expression profiling. In order to establish a gene expression index in the rice cultivar indica, we sequenced and analyzed 86,136 ESTs from nine rice cDNA libraries from the super hybrid cultivar LYP9 and its parental cultivars. We assembled these ESTs into 13,232 contigs and leave 8,976 singletons. Overall, 7,497 sequences were found similar to the existing sequences in GenBank and 14,711 are novel. These sequences are classified by molecular function, biological process and pathways according to the Gene Ontology. We compared our sequenced ESTs with the publicly available 95,000 ESTs from japonica, and found little sequence variation, despite the large difference between genome sequences. We then assembled the combined 173,000 rice ESTs for further analysis. Using the pooled ESTs, we compared gene expression in metabolism pathway between rice and Arabidopsis according to KEGG. We further profiled gene expression patterns in different tissues, developmental stages, and in a conditional sterile mutant, after checking the libraries are comparable by means of sequence coverage. We also identified some possible library specific genes and a number of enzymes and transcription factors that contribute to rice development.

An Experimental Study of Turbulent Flow in Attachment Jet Combustors by LDV

Flame stabilization in attachment jet combustors is based on the existence of the high temperature recirculation zone,provided by the Coanda effect of an attachment jet.The single attachment jet in a rectangular channel is a fundamental form of this type of flow.In this paper,the detailed characteristics of turbulent flow of a single attachment jet were experimentally studied by using a 2-D LDV,The flowfield consists of a forward flow and two reverse flows.The forward one is composed of a curved and a straight section.The curved section resembles a bent turbulent free jet,and the straight part is basically a section of turbulent wall jet.A turbulent couter-gradient transport region exists at the curved section.According to the results,this kind of combustor should have a large sudden enlarge ment ratio and not too narrow in width.