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ABSTRACT: The expression of phosphatase and tensin homolog (PTEN), a tumor suppressor gene, is frequently down-regulated in gastric carcinomas due to mutation, loss of heterozygosity, and promoter hypermethylation. However, it is unknown if additional mechanisms may account for the down-regulation of PTEN expression. While neuronal precursor cell-expressed developmentally down-regulated 4-1 (NEDD4-1) is believed to be a potential dual regulator of PTEN, there are conflicting reports regarding their interaction. To gain further insight into the role of NEDD4-1 and its association with PTEN in gastric carcinoma development, we measured the protein expression of NEDD4-1 and PTEN in gastric mucosae with various pathological lesions and found that NEDD4-1 increased from normal gastric mucosa to intestinal metaplasia and decreased from dysplasia to gastric carcinoma. These changes did not correlate with PTEN expression changes during gastric carcinogenesis. Moreover, we found similar results in protein levels in the primary tumors and adjacent non-tumorous tissues. These results differ from a previous report showing that expression of NEDD4-1 is up-regulated in gastric carcinomas, and show a more complex pattern of NEDD4-1 gene expression during gastric carcinogenesis.
World Journal of Gastroenterology 11/2012; 18(43):6345-8. · 2.47 Impact Factor
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ABSTRACT: AIM: PTEN is a tumor suppressor gene in different cancers. This study was to determine the protein expression of PTEN and phosphorylation of PTEN (p-PTEN) at residue Ser380 in different histology specimens of gastric tissues. METHODS: A total of 179tissue specimens of normal gastric mucosa, chronic gastritis, intestinal metaplasia, dysplasia, and gastric cancer were recruited for immunohistochemical analysis of PTEN and p-PTEN expression. Four gastric cancer AGS, MKN-45, MKN-28, and SGC-7901 cell lines and a non-cancerous gastric GES-1 cell line were used to detect expression of PTEN and p-PTEN protein using Western blot. RESULTS: Expression level of PTEN protein was significantly decreased in gastric cancer tissues compared to normal gastric mucosa, chronic gastritis, intestinal metaplasia and dysplasia (P<0.05). In contrast, p-PTEN protein level was significantly increased in intestinal metaplasia, dysplasia and gastric cancer compared to normal gastric mucosa and chronic gastritis (P<0.05). However, there was no any association of PTEN and p-PTEN expression with clinicopathological characteristics from gastric cancer patients. Moreover, the ratio of p-PTEN and PTEN was higher in gastric cancer cell lines than that of the non-malignant cells. CONCLUSIONS: This study demonstrated that aberrant expression of PTEN and p-PTEN at residue Ser380 was early event that could contribute to gastric carcinogenesis, and that PTEN phosphorylation at residue Ser380 could be a mechanism for PTEN inactivation.
Gastroentérologie Clinique et Biologique 04/2012; · 0.80 Impact Factor
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ABSTRACT: To explore the association of p53-MDM2 feedback loop related proteins with gastric pathologies in relation to Helicobacter pylori infection.
Gastric biopsies were obtained from 157 H. pylori-negative and positive patients, including normal gastric mucosa (NGM), chronic gastritis (CG), intestinal metaplasia (IM), dysplasia (Dys), and gastric cancer (GC). The expression of mutant p53, MDM2, Bax and PUMA in gastric tissues was detected by immunohistochemistry.
Overall expression of MDM2 and Bax is progressively increased from NGM to GC. PUMA expression is increased in CG but subsequently decreased after the development of IM. H. pylori infection is associated with increased mutant p53 and Bax expression but decreased PUMA expression in IM, and increased MDM2 expression in Dys.
These results suggest that different p53-MDM2 feedback loop related proteins are distinctly expressed in the various stages of gastric carcinogenesis; their roles in gastric carcinogenesis in the presence of H. pylori infection need to be further investigated.
Gastroentérologie Clinique et Biologique 02/2012; 36(3):235-43. · 0.80 Impact Factor
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ABSTRACT: To explore the immunity modulation function of aqueous of Forsythia suspense (AFS) and its possible mechanisms.
Rats of burned model group were burned with vapor under 3 mpa pressure and 108 degrees C temperature for 8 seconds to achieve deep partial-thickness burn, to make a thirty percent total body surface area (TBSA) burn. The experiment were divided into five groups: Control group: without any treatment; 8PBH group: 8 h after burn; the rats of AFS1 group, AFS2 group and AFS3 group of them were given AFS 5 g/kg, 2.5 g/kg, 1.25 g/kg once a day by Po. pathway for seven days before burns, respectively. Rats were sacrificed before and 8h after burn, The percentage of Treg cells in CD4(+) T cells was detected by flow cytometry (FCM); the expression of Foxp3 mRNA on splenocytes were measured by RT-PCR, and the protein of Foxp3 activity was evaluated by immunohistochemistry staining.
Compared with Control group, the expression of Foxp3 mRNA and protein on the splenocytes were upregulated markedly (P<0.01), and the percentage of Treg were significantly increased (P<0.01) in the 8PBH group. AFS1, AFS2 and AFS3 significantly attenuated these increases (P<0.01), which was dose-dependent.
AFS has immunity modulation function and mechanism of it is corrected with Foxp3 mRNA on splenocytes.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 10/2009; 25(10):935-7.
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ABSTRACT: To investigate the effect of ursolic acid on proliferation and apoptosis of hepatic stellate cells (HSC) in vitro and explore the mechanisms of apoptosis of HSC induced by ursolic acid by studying the expressions of apoptosis-regulating proteins Bcl-2, Bax and Caspase 3 in HSC.
Hepatic stellate cells HSC-T6 and hepatocytes L02 were incubated with different concentrations of ursolic acid (25, 50, 75, 100, 125 and 150 micromol/L) for 24 h, 48 h and 72 h. The effect of ursolic acid on the cell proliferation was studied by methyl thiazolyl tetrazolium (MTT) colorimetric assay. The rate of HSC-T6 apoptosis was identified by flow cytometry (FCM) and the morphological change of apoptosis was observed with light microscopy. The expressions of apoptosis-regulating protein Bcl-2, Bax and Caspase 3 in HSC-T6 after apoptosis induced by ursolic acid were examined by immunocytochemical staining assay.
MTT analysis indicated administration of 25-150 micromol/L ursolic acid incubated with HSC-T6 for 24 h, 48 h and 72 h significantly inhibited HSC-T6 proliferation in a dose-dependent and time-dependent manner compared with the control group. Promotive effect of ursolic acid on proliferation of hepatocyte L02 was observed in the 25, 50, 75 micromol/L concentration groups. Ursolic acid inhibited L02 proliferation when its concentration was higher than 100 micromol/L and for 72 hours or longer. HE stained histological slides demonstrated morphologic changes of HSC-T6, including karyorrhexis and cytoplasm vacuolization, when they were treated with ursolic acid at 75 micromol/L concentrations for 48 h. FCM showed the apoptosis ratios of HSC-T6 were 10.30%+/-3.85%, 21.87%+/-4.46% and 31.33%+/-6.18% after treating HSC-T6 with ursolic acid at concentrations of 25, 50 and 75 micromol/L for 48 h. They were significantly higher than that of the control group 2.93%+/-1.60%. Immunocytochemistry also indicated the expressions of Bax and caspase 3 protein in HSC-T6 cells were up-regulated in a dose-dependent manner, but expressions of Bcl-2 protein were not significantly different from that of the blank control group (P more than 0.05).
Ursolic acid could significantly inhibit HSC proliferation and induce apoptosis in a dose-dependent and time-dependent manner. Ursolic acid in low concentration promotes proliferation of L02 cells, but in high concentrations (more than 100 micromol/L) it inhibits the growth of hepatocytes. Expressions of Bax and Caspase 3 in apoptotic HSC were increased; expressions of Bcl-2 protein were not significantly different from that of the control group, while Bcl-2/Bax ratio was reduced. Our results suggest that HSC-T6 cell apoptosis induced by ursolic acid occurs through mechanisms involving mitochondrial pathways and Bcl-2 family proteins.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology 05/2008; 16(4):298-301.
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ABSTRACT: To investigate the influence of heat shock (HS) preconditioning on the ultra-structure of gastric mucosal cells and the activity of cytochrome oxidase (CCO) and superoxide dismutase (SOD) in mitochondria isolated from gastric mucosa of burned rats in order to investigate its protective mechanism on burn-induced acute gastric mucosal lesion.
Ninety-six Wistar rats were divided randomly into two groups:(1) burn model group (n =40) undergoing burning and made into model of acute gastric mucosal lesion; and then subdivided into 5 equal subgroups: 3, 6, 12, 24, and 48 h after burning; (2) normal control group (n = 8); (3) HS preconditioning + burn group (n = 40), undergoing burning 24 h after HS preconditioning; and (4) HS preconditioning group (n = 8), without burn as experimental control A number of rats were sacrificed and laparotomized before and 3, 6, 12, 24, and 48 h after the burning. Specimens were obtained from the gastric antrum, bodies if stomach and gastric lesion to undergo microscopy and determination of the ulcer index ( UI ). Streptavidin-peroxidase immunohistochemistry was used to detect the protein expression of heat shock protein (HSP)-60 and HSP-70. Mitochondria were isolated and the activities of CCO and SOD in mitochondria were measured. The dynamic postburn changes in micro and ultra structure of gastric mucosal cells of scalded rats were observed.
The rats of the burn group showed conspicuous gastric mucosal lesions. The UI levels at every time point of the HS group were all the lowest in comparison with other groups. The expression levels of HSP70 and HSP60 of the HS group, especially those at the time points 3, 6, 12, and 24 h were significantly higher than those of the burn group (P <0.05 or P <0.01). The activities of mCCO and mSOD of the HS group did not decrease remarkably at any time point in comparison with the control groups, and the activities of mCCO and mSOD at the time points 6, 12, and 24 h of the HS group were all significantly higher than those of the burn group (P <0.05 of P <0.01). Electron microscopy showed that the ultrastructural lesion was mild and alteration of gastric mucosal mitochondria was not significant in the HS group compared with those of the burn group. A positive correlation was shown between SOD and HSP70 (r = 0.436, P <0.05) and also between CCO and HSP60 (r =0.679, P <0.05).
HS preconditioning ameliorates the burn-induced acute gastric mucosal lesion and has a protective role to gastric mucosal mitochondria. The role of HS preconditioning on mitochondria is associated with the protection of the activity of antioxidase and CCO by HSP60 and 70.
Zhonghua yi xue za zhi 02/2008; 88(8):564-7.
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ABSTRACT: We consider the problem of learning an acyclic discrete circuit with n wires, fan-in bounded by k and alphabet size s using value injection queries. For the class of tran-sitively reduced circuits, we develop the Distinguishing Paths Algorithm, that learns such a circuit using (ns) O(k) value injection queries and time polynomial in the number of queries. We describe a generalization of the algorithm to the class of circuits with shortcut width bounded by b that uses (ns) O(k+b) value injection queries. Both algorithms use value injec-tion queries that fix only O(kd) wires, where d is the depth of the target circuit. We give a reduction showing that without such restrictions on the topology of the circuit, the learn-ing problem may be computationally intractable when s = n Θ(1) , even for circuits of depth O(log n). We then apply our large-alphabet learning algorithms to the problem of approxi-mate learning of analog circuits whose gate functions satisfy a Lipschitz condition. Finally, we consider models in which behavioral equivalence queries are also available, and extend and improve the learning algorithms of (Angluin in Proceedings of the Thirty-Eighth Annual ACM Symposium on Theory of Computing, pp. 584–593, 2006) to handle general classes of gate functions that are polynomial time learnable from counterexamples.
Machine Learning 01/2008; 72(72):113-138. · 1.59 Impact Factor
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ABSTRACT: To study the cellular immunity induced by H.pylori vaccine with chitosan as adjuvant and the mechanism of immunological protection.
BALB/c mice were randomly divided into nine groups and immunized by (1)PBS alone, (2)chitosan solution alone, (3)chitosan particles alone, (4)H.pylori antigen alone, (5)H.pylori antigen plus chitosan solution, (6)H.pylori antigen plus chitosan particles, (7)H.pylori antigen plus cholera toxin (CT), (8)H.pylori antigen plus chitosan solution and CT, (9)H.pylori antigen plus chitosan particles and CT orally respectively once a week for four weeks. At 4 weeks after the last immunization, mice were challenged by alive H.pylori (1x10(9) CFU/mL) twice at two days intervals. Before and after the challenge, mice were killed in batches and the H.pylori-infection in gastric mucosa was detected by H.pylori culture and Giemsa stain. ELISA and HE stain were used to detect IL-2, IL-4, IL-10 levels and pathologic change in gastric mucosa.
(1)In the groups with chitosan as an adjuvant, 60% mice could achieve immunological protection, which was consistent with using CT as an adjuvant (58.33%), and was more than that when using H.pylori antigen alone or without H.pylori antigen. (2)After challenge, the IL-2 levels in gastric mucosa in the groups with adjuvants were significantly higher than those in the control group (P<0.001-0.05). Moreover, IL-2 levels in the groups with adjuvants after challenge were significantly higher than those before challenge (P<0.05). Before challenge, the IL-10 and IL-4 levels in gastric mucosa were significantly higher in the groups with chitosan as adjuvant than those in non-adjuvant groups (P<0.05). After challenge, IL-10 levels were significantly higher in the groups with chitosan particles as adjuvant than those in other groups (P<0.05); IL-4 levels were significantly higher in the groups with chitosan particles as an adjuvant than those in the group with CT as an adjuvant, and those in the group with chitosan solution as an adjuvant were significantly higher than those in control group, non-adjuvant group and the groups with CT (P<0.05). IL-10 and IL-4 in the groups with adjuvants after challenge were significantly lower than those before challenge (P<0.05). (3)The degree of inflammation in gastric mucosa was significantly lower in the groups with chitosan and chitosan particles as adjuvant than those with CT as adjuvant(P<0.05).
(1)H.pylori vaccine with chitosan as an adjuvant has the immune protective effect against H.pylori infection. (2)H.pylori vaccine with chitosan as an adjuvant could reverse the inhibition of Th2 induced by H.pylori infection and recover the Th1/Th2 imbalance, which might contribute to the immune protection against H.pylori. (3)The rate of gastritis induced by H.pylori vaccine with chitosan as adjuvant was significantly lower than those with CT as adjuvant.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 08/2007; 23(7):595-9.
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ABSTRACT: To study the anti-Helicobacter pylori (Hp) effect and the regulation of T helper (Th) response of chitosan.
Hp infected grade one female BALB/c mice model was established by inoculating Hp Sydney strain 1 and the mice were randomly divided into eight groups and administrated with (1) Arabia glue solution (control group), (2) omeprazole, (3) amoxicillin, (4) amoxicillin plus omeprazole, (5) chitosan, (6) chitosan plus omeprazole, (7) chitosan plus amoxicillin, (8) chitosan plus amoxicillin plus omeprazole respectively twice daily for 14 consecutive days. Four weeks after the last administration, these mice were all killed and samples of gastric mucosa were embedded in paraffin, sectioned and assayed with Giemsa stain. The remaining gastric mucosa was used to quantitatively culture Hp. An quantitative ELISA was used to detect IL-2, IFNgamma, IL-12, IL-4, IL-10 content in gastric mucosa.
The eradication rate of Hp was 0, 0, 41.7%, 58.3%, 58.3%, 66.7%, 83.3% and 91.7% respectively among these eight groups (P < 0.01). The Hp colony density in the (1) group and (2) group was significantly higher than that in the other six groups (P < 0.05). The Hp colony density in (3) group was significantly higher than that in (6) group, (7) group and (8) group (P < 0.05) and that in (4) group was significantly higher than that in (8) group (P < 0.05). There was no difference in the content of IL-2 in the gastric mucosa among these eight groups (P > 0.05). The content of IFNgamma, IL-12, IL-4 and IL-10 in the gastric mucosa in groups with chitosan was significantly higher than that in groups without chitosan (P < 0.05).
Chitosan has anti-Hp effect and synergism with amoxicillin in vivo. Chitosan can up-regulate Th1 and Th2 response.
Zhonghua nei ke za zhi [Chinese journal of internal medicine] 04/2007; 46(3):220-3.
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ABSTRACT: To determine the effect of pioglitazone, a specific peroxisome proliferator-activated receptor-gamma (PPARgamma) ligand, on development of severe acute pancreatitis (SAP) and expression of nuclear factor-kappa B (NF-kappaB) and intercellular adhesion molecule-1 (ICAM-1) in the pancreas.
Male Sprague-Dawley (SD) rats (160-200 g) were randomly allocated into three groups (n = 18 in each group): severe acute pancreatitis group, pioglitazone group, sham group. SAP was induced by retrograde infusion of 1 mL/kg body weight 5% sodium taurocholate (STC) into the biliopancreatic duct of male SD rats. Pioglitazone was injected intraperitoneally two hours piror to STC infusion. Blood and ascites were obtained for detecting amylase and ascitic capacity. Pancreatic wet/dry weight ratio, expression of NF-kappaB and ICAM-1 in pancreatic tissues were detected by immunohistochemical staining. Pancreatic tissue samples were stained with hematoxylin and eosin (HE) for routine optic microscopy.
Sham group displayed normal pancreatic structure. SAP group showed diffuse hemorrhage, necrosis and severe edema in focal areas of pancreas. There was obvious adipo-saponification in abdominal cavity. Characteristics such as pancreatic hemorrhage, necrosis, severe edema and adipo-saponification were found in pioglitazone group, but the levels of those injuries were lower in pioglitazone group than those in SAP group. The wet/dry pancreatic weight ratio, ascetic capacity, serum and ascitic activities of anylase in the SAP group were significantly higher than those in the sham group and pioglitazone group respectively (6969.50 +/- 1368.99 vs 2104.67 +/- 377.16, 3.99 +/- 1.22 vs 2.48 +/- 0.74, P < 0.01 or P < 0.05). According to Kusske criteria, the pancreatic histologic score showed that interstitial edema, inflammatory infiltration, parenchyma necrosis and parenchyma hommorrhage in SAP group significantly differed from those in the sham group and pioglitazone group (7.17 +/- 1.83 vs 0.50 +/- 0.55, 7.67 +/- 0.82 vs 6.83 +/- 0.75, P < 0.01, P < 0.05. The expression of NF-kappaB and ICAM-1 in sham group was lower than that in SAP group and pioglitazone group (0.50 +/- 0.55 vs 33 +/- 1.21, P < 0.01). There was a significant difference in the expression of NF-kappaB and ICAM-1 between SAP group and pioglitazone group (7.50 +/- 1.05 vs 11.33 +/- 1.75, 0.80 +/- 0.53 vs 1.36 +/- 0.54, P < 0.01 or P < 0.05) at 12 h after the induction of pancreatitis.
Pioglitazone attenuates the severity of SAP. The beneficial effect of pioglitazone is multifactorial due to its anti-inflammatory activities, most likely through the inhibition of ICAM-1 expression and NF-kappaB activation. Specific ligands of PPARgamma may represent the novel and effective means of clinical therapy for SAP.
World Journal of Gastroenterology 04/2007; 13(13):1983-8. · 2.47 Impact Factor
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ABSTRACT: To study the immunological protective effect of H pylori vaccine with chitosan as an adjuvant and its mechanism.
Female BALB/c mice were randomly divided into seven groups and orally immunized respectively with PBS, chitosan solution, chitosan particles, H pylori antigen, H pylori antigen plus cholera toxin (CT), H pylori antigen plus chitosan solution, H pylori antigen plus chitosan particles once a week for four weeks. Four weeks after the last immunization, the mice were challenged twice by alive H pylori (1 x 10(9) CFU/mL) and sacrificed. Part of the gastric mucosa was embedded in paraffin, cut into sections and assayed with Giemsa staining. Part of the gastric mucosa was used to quantitatively culture H pylori. ELISA was used to detect cytokine level in gastric mucosa and anti- H pylori IgG1, IgG2a levels in serum.
In the groups with chitosan as an adjuvant, immunological protection was achieved in 60% mice, which was significantly higher than in groups with H pylori antigen alone and without H pylori antigen (P < 0.05 or 0.001). Before challenge, the level of IFN and IL-12 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in the control group and the group without adjuvant (P < 0.05 or 0.005). After challenge, the level of IFN and IL-12 was significantly higher in the groups with adjuvant than in the groups without adjuvant and antigen (P < 0.05 or 0.001). Before challenge, the level of IL-2 in gastric mucosa was not different among different groups. After challenge the level of IL-2 was significantly higher in the groups with adjuvant than in the control group (P < 0.05 or 0.001). Before challenge, the level of IL-10 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in other groups without adjuvant (P < 0.05 or 0.01). After challenge, the level of IL-10 was not different among different groups. Before challenge, the level of IL-4 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in other groups without adjuvant (P < 0.05). After challenge, the level of IL-4 was significantly higher in the groups with chitosan particles as an adjuvant than in the group with CT as an adjuvant (P < 0.05), and in the group with chitosan solution as an adjuvant, the level of IL-4 was significantly higher than that in control group, non-adjuvant group and the groups with CT (P < 0.05 or 0.001). The ratio of anti- H pylori IgG2a/IgG1 in serum was significantly lower in the groups with chitosan as an adjuvant than in the groups with CT as an adjuvant or without adjuvant (P < 0.01).
H pylori vaccine with chitosan as an adjuvant can protect against H pylori infection and induce both Th1 and Th2 type immune response.
World Journal of Gastroenterology 03/2007; 13(10):1547-53. · 2.47 Impact Factor
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ABSTRACT: To study the endoscopic, pathological and immunohistochemical features of esophageal mesenchymal tumors.
Twenty-nine patients diagnosed as esophageal mesenchymal tumors by electronic endoscopy and endoscopic ultrasound (EUS) were observed under light microscopes, and all tissues were stained by the immunohistochemical method. The expression of CD117, CD34, SMA and desmin were measured by staining intensity of cells and positive cell ratios.
Endoscopically, esophageal gastrointestinal stromal tumors (GISTs) and leiomyomas (LMs) had similar appearances, showing submucosal protuberant lesions. They all showed low echo images originated from the muscularis propria or muscularis mucosa on EUS. Endoscopy and EUS could not exactly differentiate esophageal GISTs from LMs. Microscopically, there were two kinds of cells: spindle cell type and epitheloid cell type in esophageal GISTs. Leiomyomas and leiomyosarcomas were only of spindle cell type. One malignancy was found in five cases of esophageal GISTs, and one malignancy in 24 cases of leiomyomas and leiomyosarcomas. Using Fisheros exact method, the differences of malignant lesion proportion were not significant between esophageal LMs and GISTs, 1/5 vs 1/24 (P > 0.05). All cases of esophageal GISTs were positive for CD117, and 3 cases were also positive for CD34. The 24 cases of leiomyomas and leiomyosarcomas were all negative for CD117 and CD34. The differences of positive rates of CD117 and CD34 were significant between esophageal GISTs and LMs, 5/5 vs 0/24, 3/5 vs 0/24 (P < 0.005). All leiomyomas and leiomyosarcomas were positive for SMA, and desmin. Among 5 cases of esophageal GISTs, 2 cases were SMA positive, and 1 case was desmin positive. The differences in positive rates and expression intensity of SMA and desmin were significant between esophageal LMs and GISTs, 24/24 vs 2/5, 24/24 vs 1/5 (P < 0.005).
The most common esophageal mesenchymal tumors are leiomyomas, and esophageal GISTs are less common. Most of esophageal LMs and GISTs are benign. Endoscopy and EUS are the effective methods to diagnose esophageal mesenchymal tumors and they can provide useful information for the treatment of these tumors. However, they cannot exactly differentiate esophageal GISTs from LMs. Pathological, especially immunohistochemical features are useful to differentiate GISTs from leiomyomas.
World Journal of Gastroenterology 03/2007; 13(5):768-73. · 2.47 Impact Factor
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ABSTRACT: To observe the influence of heat shock preconditioning on the expressions of heat shock protein (HSP) 60 and HSP 70 and on the activities of cytochrome oxidase (CCO) and superoxide dismutase (SOD) in mitochondria in gastric mucosa of severely scalded rats, and to investigate its protective mechanism on acute gastric mucosal lesion in rats with severe scald.
One hundred and forty-four Wistar rats were randomized into three groups, i. e. scald group ( n = 40, acute gastric mucosal lesion was made after scald, other 8 normal rats without scald were employed as blank control); HS group ( n =40, with heat shock preconditioning 20 h before scald), and other 8 rats preconditioned with heat shock but without scald were employed as experimental control I; actinomycin D group ( n = 40, with intraperitoneal injection of 0. 1 mg/kg actinomycin D 30 min before heat shock preconditioning and other treatment as HS group), and other 8 rats with merely actinomycin D injection were employed as experimental control II. Eight rats in each group were sacrificed and laparotomized at 3, 6, 12, 24 and 48 post-scald hours (PSH) , respectively to determine the index of gastric mucosal lesions (UI ) , the mRNA expressions of HSP70 and protein expression of HSP60 and HSP70, and the changes in the activities of SOD and CCO.
UI of the scalded rats increased as the time elapses, reaching the peak (12. 8 +/- 1.9) at 24 PSH. In addition, UI in HS group was significantly lower than that in scald group at each time-point except that at 3 PSH ( P < 0. 05 or 0. 01). The extent of gastric mucosal lesion in rats in actinomycin D group was obviously aggravated compared with that in scald and HS groups ( P <0. 05). The HSP70 mRNA expression in both scald and HS groups was increased at each time-points except for 48PBH, while that in actinomycin D group was increased at 24 PBH and 48PBH. The expressions of HSP70 and HSP60 were greatly increased in HS group compared with those in scald group ( P < 0. 05 or 0. 01) , while those in actinomycin D group were significantly inhibited ( P < 0. 05). The activities of CCO and SOD were gradually decreased in gastric mucosa in scald group, but it was greatly improved by HS preconditioning at 6, 12, 24 PSH ( P < 0. 05 or 0. 01).
Heat shock preconditioning is beneficial for the protection of acute gastric mucosal lesion of rats after severe scald, due to increase of HPS60 and HSP70 expression, and increase of CCO and SOD activities in mitochondria.
Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns 02/2007; 23(1):58-61.
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ABSTRACT: To evaluate the diagnostic values of the detection of mRNAs of GA733 gene family in exfoliated cells of pleural and peritoneal effusions.
Sixty specimens of pleural and peritoneal fluids from 60 patients were collected. GA733-1 and GA733-2 mRNA in the exfoliated cells were detected by qualitative RT-PCR, and their diagnostic values were assessed. Patients Sixty patients with pleural or peritoneal effusions, from May 2003 and August 2004, aged 23-85 (average 56.5 years).
GA733-1 and GA733-2 mRNA were positive in 5 (13.9%) and 27 (75.0%) of 36 malignant specimens, and in 1 and 7 of 11 cause-unknown specimens, respectively, but both of them were negative in all 13 benign specimens, and the difference of GA733-2 mRNA positive rates among the three groups was significant (P<0.005), but that of GA733-1 mRNA was not (P>0.05). GA733-1 and GA733-2 mRNA were positive in 4 (15.4%) and 22 (84.6%) of 26 cytological positive specimens, and in 1 and 6 of 6 suspicious specimens, and in 1 (3.5%) and 6 (21.4%) of 28 negative specimens, respectively and the difference of GA733-2 mRNA positive rates among the three groups was significant (P<0.005), but that of GA733-1 mRNA was not (P>0.05). Sensitivity, specificity and accuracy of detection for GA733-2 mRNA for diagnosis of malignant effusions were 75.0%, 100% and 81.6%, respectively.
The detection of GA733-2 mRNA by qualitative RT-PCR is sensitive and highly specific for the diagnosis of malignant pleural and peritoneal effusions, while the diagnostic value of GA733-1 mRNA needs to be further investigated.
Internal Medicine 01/2007; 46(18):1489-94. · 0.94 Impact Factor
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ABSTRACT: To investigate the expression of peroxisome proliferators-activated receptor-gamma (PPAR-gamma) and cyclooxygenase-2 (COX-2) in Helicobacter pylori (Hp)-associated gastric mucosal lesions and the relations among them.
Two-step immunohistochemical staining was used to detect the expression of PPAR-gamma and COX-2 proteins in the gastric biopsy specimens from 209 patients with lesions of gastric mucosa, including 75 cases of chronic superficial gastritis (CSG), 24 cases of chronic atrophic gastritis (CAG), and 90 cases of intestinal metaplasia (MI) and dysplasia (DYS), and from 20 subjects with normal gastric mucosa negative in Hp as controls. In addition, 36 CSG patients were followed up for 10 years to observe the changes of expression of PPAR-gamma and COX-2.
(1) The expression of COX-2 protein in the glandular cells of the CSG, CAG, and IM + DYS groups were 42.7%, 45.8%, and 61.1% respectively, all significantly higher than that of the control group (20%, P < 0.01 or 0.05). The expression of COX-2 protein in the inflammatory cells of the CSG, CAG, and IM + DYS groups were 32%, 29.2%, and 36.7% respectively, all significantly higher than that of the control group (0%, all P < 0.05). (2) In the CSG and IM + DYS groups the expression levels of COX-2 in both the inflammatory and glandular cells were both significantly higher in the Hp-positive patients than in the Hp-negative patients (P < 0.05 or 0.01). (3) The expression of PPAR-gamma protein in the glandular cells of the CSG, CAG, and IM + DYS groups were 18.7%, 29.2%, and 45.6% respectively, all significantly higher than that of the control group (0%, all P < 0.01); and the expression levels of PPAR-gamma protein in the inflammatory cells of the CSG, CAG, and IM + DYS groups were 22.5%, 20.8%, and 22.2% respectively, all significantly higher than that of the control group (0%, P < 0.05 or 0.01). (4) In the CSG and IM + DYS groups the expression levels of PPAR-gamma in the inflammatory cells of the Hp-positive patients were both significantly higher than those of the Hp-negative patients (both P < 0.01); and the. expression levels of PPAR-gamma in the glandular cells of the Hp-positive patients were both significantly higher than those of the Hp-negative patients (both P < 0.01). (5) The expression level of COX-2 in the inflammatory cells of the CSG patients with a persistent Hp infection after a period of ten years was 33.3%, significantly lower than that a decade before (38.9%, P < 0.05). However, the expression level of PPAR-gamma in the inflammatory cells of the patients with a persistent Hp infection after a period of ten years was 30.6%, not significantly different from that a decade before (33.3%, P > 0.05). (6) The expression levels of COX-2 and PPAR-gamma in the glandular cells of the CSG patients with a persistent Hp infection after a period of ten years were 55.6% and 30.6% respectively, both significantly higher than those a decade before (47.2% and 22.2%, both P < 0.01). (7) In all Hp-negative disease groups the expression level of COX-2 in the inflammatory cells was not significantly correlated with the expression level of PPAR-gamma in the inflammatory cells(r = 0.006 and 0.149, P > 0.05) In the Hp-negative disease groups, the expression level of COX-2 in the glandular cells was significantly positively correlated with the expression level of PPAR-gamma in the glandular cells in the IM + DYS group (r = 0.336, P < 0.05), however, not significantly correlated with the expression level of PPAR-gamma in the glandular cells in both CAG and CSG groups (r = 0.035 and 0.126, both P > 0.05). (8) In both the inflammatory cells and glandular cells the COX-2 expression was significantly positively correlated with the PAR-gamma expression in all disease groups with Hp positivity (r = 0.348 and 0.645, P < 0.05 or 0.01).
(1) Hp infection induces the overexpression of COX-2 and PPAR-gamma in the inflammatory and glandular cells within gastric mucosa, thus participating in the pathogenesis of gastric cancer (2) The expression of COX-2 induces by Hp is related to acute inflammation caused by Hp infection. In the glandular cells, significant increase of COX-2 and PPAR-gamma expression implies that along with time the expression of COX-2 and PPAR-gamma may change in advance of the morphological changes of the gastric mucosa.
Zhonghua yi xue za zhi 11/2006; 86(38):2683-9.
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ABSTRACT: To investigate the changes in cellular apoptosis of Peyer's patches in severely scalded mice, and to explore its role in the pathogenesis of gut barrier damage.
Forty BALB/c mice were randomly divided into normal control, 12 post-scald hour (12PSH), 24PSH and 72PSH groups, with 10 in each group. The mice in all PSH groups were inflicted with 20% TBSA full-thickness scald on the back. The mice in all the groups were sacrificed at different time points, and Peyer's patches were harvested from all the mice for HE staining, DNA gel electrophoresis, and flow cytometry ( FCM) examination with FITC conjugated Annexin-v and propidium iodide( PI) staining of cells.
HE staining revealed that there were relatively abundant apoptotic cells scattering in Peyer's patches of scalded mice . DNA electrophoresis of Peyer's patches revealed typical " ladder" pattern at all indicated time points in scalded mice. Apoptotic percentage of detached Peyer's patches cells in control and scalded group were (4. 9+/-2. 1)% , (26.7+/-3. 1)% , (21.6 +/-4.0)% ,(12. 8 +/-2.0)% , respectively, and the percentage reached the peak at 12 PSH.
Apoptosis is a principle modality of cell death of small intestinal Peyer's patches lymphocytes in severely scalded mice, and it might contribute to immunity barrier failure of intestinal wall after severe thermal injury.
Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns 09/2006; 22(4):254-7.
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ABSTRACT: This paper considers the pure bending problem of simply supported transversely isotropic circular plates with elastic compliance coefficients being arbitrary functions of the thickness coordinate. First, the partial differential equation, which is satisfied by the stress functions for the axisymmetric deformation problem is derived. Then, stress functions are obtained by proper manipulation. The analytical expressions of axial force, bending moment and displacements are then deduced through integration. And then, stress functions are employed to solve problems of transversely isotropic functionally graded circular plate, with the integral constants completely determined from boundary conditions. An elasticity solution for pure bending problem, which coincides with the available solution when degenerated into the elasticity solutions for homogenous circular plate, is thus obtained. A numerical example is finally presented to show the effect of material inhomogeneity on the elastic field in a simply supported circular plate of transversely isotropic functionally graded material (FGM).
J Zhejiang Univ SCIENCE A. 01/2006; 7:1324-1328.
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ABSTRACT: To explore the expressions of GST-Pi and telomerase activity in esophageal carcinoma and premalignant lesions and to investigate the value of endoscopic methylene blue (MB) and Lugol's iodine double staining.
Seventy-two patients with esophagopathy were sprayed endoscopically with MB and Lugol's iodine in proper order and the areas stained blue and brown, and the area between the blue and brown stains were obtained. Depending on the pattern of mucosal staining, biopsy specimen was obtained. GST-Pi and telomerase activity in specimens were examined by immunohistochemistry and PCR-based silver staining telomeric repeat amplification protocol, respectively.
After MB and Lugol's iodine staining, the area between both the colors was obtained in 64 of the 72 patients and the areas were stained blue and brown in all of the 72 patients. Association test of two simultaneous ordinal categorical data showed a correlation between the esophageal mucosal staining and the esophageal histology (P<0.005). The expression of GST-Pi and telomerase activity in esophageal carcinoma and premalignant lesions increased. The expression of GST-Pi and telomerase activity in dysplasia and carcinoma was significantly higher than that in normal epithelium (P<0.005). The expression in hyperplasia was slightly higher than that in normal epithelium. With the lesions progressing from low- to moderate- to high-grade dysplasia, the positive rate increased (P<0.025). Expression of GST-Pi was correlated with that of telomerase activity in dysplasia and carcinoma (phis = 0.4831, P<0.005; phis = 0.3031, P<0.025, respectively); but there was no correlation between them in normal epithelium and hyperplasia.
The expression of GST-Pi and telomerase may be an early event in the carcinogenesis of esophagus. They may play an induced and synergistic role with each other in the carcinogenesis of esophagus. Endoscopic MB and Lugol's iodine double staining and detection of GST-Pi and telomerase activity may contribute to the early diagnosis of esophageal carcinoma.
World Journal of Gastroenterology 10/2005; 11(39):6090-5. · 2.47 Impact Factor
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ABSTRACT: To study the immunological protection of Helicobacter pylori (H. pylori) vaccine with chitosan as adjuvant and it's mechanism.
One-grade female BALB/c mice were randomly divided into nine groups and immunized by (1) PBS alone, (2) chitosan solution alone, (3) chitosan particles alone, (4) H. pylori antigen alone, (5) H. pylori antigen plus chitosan solution, (6) H. pylori antigen plus chitosan particles, (7) H. pylori antigen plus CT, (8) H. pylori antigen plus chitosan solution and cholera toxin (CT), (9) H. pylori antigen plus chitosan particles and CT orally respectively once a week for four weeks. At 4 weeks after the last immunization, these mice were challenged by alive H. pylori (1 x 10(9)/ml) twice at two days intervals. At 4 weeks after the last challenge, these mice were all killed and Gastric mucosa were embedded in paraffin, sectioned and assayed with Giemsa stain. The other gastric mucosa were used to quantitatively culture H. pylori. An ELISA was used to detect anti-H. pylori IgA in saliva and gastric mucosa and a quantitative ELISA was used to detect IL-2, IL-4, IL-10 content in gastric mucosa, and SP immunohistochemical method was used to detect secretory immunoglobulin A (sIgA) in gastric mucosa.
(1) In the groups with chitosan as adjuvant, 60% mice could achieve immunological protection, which was according to that with CT as adjuvant (58.33%), and was significantly higher than H. pylori antigen alone and other groups without H. pylori antigen (P < 0.01 or P < 0.05). While the rates of protection in the groups with chitosan plus CT as adjuvant were 84.62%, 85.71% and the H. pylori colonization score in it was significantly lower than that in the groups with CT as adjuvant and without adjuvants (P < 0.01 or P < 0.05). (2) the labeling index for sIgA-positive lumen of glands and special anti-H. pylori IgA levels in gastric mucosa in the groups with chitosan as an adjuvant had no difference with those in the group with CT as an adjuvant (P > 0.05) and were significantly higher than those in non-adjuvant groups, while those in the groups with chitosan plus CT were significantly higher than those in the group with CT as an adjuvant (P < 0.01 or P < 0.05). (3) Before challenge, the content of IL-2 in gastric mucosa were no different among different groups (P > 0.05). After challenge the content of IL-2 were significantly higher in the groups with adjuvant than those in the control group (P < 0.01 or P < 0.05), Moreover, those in the groups with antigen after challenge were significantly higher than those before challenge (P < 0.05). (4) Before challenge, the content of IL-10 in gastric mucosa were significantly higher in the groups with chitosan as adjuvant than those in the control group and the group without adjuvant (P < 0.01 or P < 0.05). After challenge, the content of IL-10 were no different among different groups (P > 0.05). Moreover, those in the groups with adjuvant after challenge were significantly lower than those before challenge (P < 0.01). (5) Before challenge, the content of IL-4 in gastric mucosa were significantly higher in the groups with chitosan as adjuvant than those in the control group and the group without adjuvant (P < 0.05), After challenge, the content of IL-4 were significantly higher in the groups with chitosan particles as an adjuvant than those in the group with CT as an adjuvant (P < 0.05), and those in the group with chitosan solution as an adjuvant were significantly higher than those in control group, non-adjuvant group and the groups with CT (P < 0.01 or P < 0.05), Moreover, those in the groups with adjuvant after challenge were significantly lower than those before challenge (P < 0.01 or P < 0.05).
(1) H. pylori vaccine with chitosan as adjuvant could protect against H. pylori infection, this suggested that chitosan could be a mucosa adjuvant of H. pylori vaccine, and it could effectively elicit special humoral immune response of systemic and local mucosa, which might be one of its protective mechanism. (2) H. pylori vaccine with chitosan as adjuvant may induce both Th1 and Th2 type immune response, and after challenge it could reverse the inhibition of Th2 induced by H. pylori infection and recover the Th1/Th2 imbalance. which might contribute to the immune protection against H. pylori.
Zhonghua yi xue za zhi 09/2005; 85(37):2629-35.
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ABSTRACT: To study the expression of hypoxia inducible factor-1alpha (HIF-1alpha) protein in prostate cancer (Pca) and its biological significance, the expression of HIF-1alpha was assayed by means of immunohistochemical technique in 42 prostate cancer, 12 prostatic intraepithelial neoplasm (PIN) and 9 normal prostate tissue (NP) specimens. Western blot was used to examine the expression of HIF-1alpha in prostate cancer cell line (PC-3M) induced by different oxygen tension. HIF-1alpha expression was positive in 33 Pca and 9 PIN specimens, and the positive rate of HIF-1alpha was higher in distant metastasis patients than in patients without metastasis of prostate cancer (P<0.05), while there was no expression of HIF-1alpha in NP. The level of HIF-1alpha in PC-3M significantly increased with the decrease of oxygen tension (P<0.01). Overexpression of HIF-1alpha is the preliminary event of the formation of Pca, which may induce carcinoma into malignant phenotype. Thus it may serve as an early diagnosis marker and the novel target for Pca treatment.
Journal of Huazhong University of Science and Technology 01/2004; 24(6):593-5. · 0.38 Impact Factor
