AIM: To study the effect of NF-κB, survivin, Bd-2 and Caspase3 on tumor necrosis factors related apoptosis inducing ligand (TRAIL) induced apoptosis of gastric cancer cells. METHODS: Gastric cancer cells of SGC-7901,...AIM: To study the effect of NF-κB, survivin, Bd-2 and Caspase3 on tumor necrosis factors related apoptosis inducing ligand (TRAIL) induced apoptosis of gastric cancer cells. METHODS: Gastric cancer cells of SGC-7901, MKN28, MKN45 and AGS lines were cultured in PRMI-1640 medium and the apoptosis rates of the cells of 4 lines were observed after treatment of tumor necrosis factors related apoptosis indudng ligand (TRAIL) with a flow cytometer. The expression of NF-κB, survivin, Bcl-2 and Caspase3 in gastric cancer cells of 4 lines was analyzed with Western blot. RESULTS: After the gastric cancer cells were exposed to TRAIL 300 ng/ml for 24 hours, the apoptosis rate was 36.05%, 20.27%, 16.50% and 11.80% in MKN28, MKN45,AGS and SC-C-7901cells respectively. Western blot revealed that the expressions of NF-EB and survivin were lower in MKN28 cells than in MKN45, AGS and SGC-7901 cells. In contrast, the expression of Caspase3 was higher in MKN28 cells than in MKN45, AGS and SGC-7901 cells. CONCLUSION: There is a selectivity of TRAIL potency to induce apoptosis in gastric cancer cells of different cell lines.The anticancer potency of TRAIL is associated with the decreased expression of NF-κB and survivin and increased expression of Caspase3 of gastric cancer cells.展开更多
AIM: To test the hypothesis that introduction of antisense TβR Ⅰ and TβR Ⅱ eukaryotic expressing plasmids into a rat model of immunologically induced liver fibrosis might block the action of TGF-β1 and halt the p...AIM: To test the hypothesis that introduction of antisense TβR Ⅰ and TβR Ⅱ eukaryotic expressing plasmids into a rat model of immunologically induced liver fibrosis might block the action of TGF-β1 and halt the progression of liver fibrosis. METHODS: RT-Nest-PCR and gene recombination techniques were used to construct rat antisense TβR Ⅰ and TβR Ⅱ recombinant plasmids which could be expressed in eukaryotic cells. The recombinant plasmids and empty vector (pcDNA3) were encapsulated by glycosyl-poly-L-lysine and then transducted into rats of pig serum-induced liver fibrosis model. Expression of exogenously transfected gene was assessed by Northern blot, and hepatic expressions of TβR Ⅰ and TβR Ⅱ were evaluated by RT-PCR and Western blot.We also performed ELISA for serum TGF-β1, hydroxyproline of hepatic tissues, immunohistochemistry for collagen types Ⅰ and Ⅲ, and VG staining for pathological study of the liver tissues. RESULTS: The exogenous antisense TβR Ⅰ and TβR Ⅱ plasmids could be well expressed in vivo, and block mRNA and protein expression of TβR Ⅰ and TβR Ⅱ in the fibrotic liver at the level of mRNA respectively. These exogenous plasmid expressions reduced the level of TGF-β1 (antisense TβR Ⅰ group 23.998+3.045 ng/mL, antisense TβR Ⅱ group 23.156+3.131 ng/mL, disease control group 32.960+3.789 ng/mL; F-=-38.19, 36.73, P<0.01). Compared with disease control group, the contents of hepatic hydroxyproline (antisense TβR Ⅰ group 0.169+0.015 mg/g liver, antisense TβR Ⅱ group 0.167+0.009 mg/g liver, disease control group 0.296+0.026 mg/g liver; F=14.39, 15.48, P<0.01) and the deposition of collagen types Ⅰ and Ⅲ decreased in the two antisense treatment groups(antisense TβR Ⅰ group, collagen type Ⅰ 669.90+50.67, collagen type Ⅲ 657.29+49.48; antisense TβR Ⅱ group, collagen type Ⅰ 650.26+51.51, collagen type Ⅲ 661.58+55.28; disease control group, collagen type I 1209.44+116.60, collagen type Ⅲ 1175.14+121.44; F=15.48 to 74.89, P<0.01). Their expression also improved the pathologic classification of liver fibrosis models (compared with disease control group, X^2=17.14, 17.24, P<0.01). No difference was found in the level of TGF-β1, the contents of hepatic hydroxyproline and collagen types Ⅰ and Ⅲ and pathologic grade between pcDNA3 control group and disease control group or between the two antisense treatment groups (F=0.11 to1.06, X^2=0.13 to 0.16, P>0.05). CONCLUSION: Antisense TβR Ⅰ and TβR Ⅱ recombinant plasmids have certain reverse effects on liver fibrosis and can be used as possible candidates for gene therapy.展开更多
AIM: To investigate the effects of platelet-derived growth factor(PDGF) and interleukin-10 (IL-10) on Fas/Fas-ligand and Bcl-2/Bax mRNA expressions in rat hepatic stellate cells.METHODS: Rat hepatic stellate cells (HS...AIM: To investigate the effects of platelet-derived growth factor(PDGF) and interleukin-10 (IL-10) on Fas/Fas-ligand and Bcl-2/Bax mRNA expressions in rat hepatic stellate cells.METHODS: Rat hepatic stellate cells (HSCs) were isolated and purified from rat liver by in situ digestion of collagenase and pronase and single-step density Nycodenz gradient.After activated by culture in vitro, HSCs were divided into 4 groups and treated with nothing (group N), PDGF (group P),IL-10 (group I) and PDGF in combinalJon with IL-10 (group C),respectively. Semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis was employed to compare the mRNA expression levels of Fas/FasL and Bcl-2/Bax in HSCs of each group.RESULTS: The expression levels of Fas between the 4 groups had no significant differences (P>0.05). FasL mRNA level in normal culture-activated HSCs (group N) was very low.It increased obviously after HSCs were treated with IL-10(group I) (0.091±0.007 vs 0.385±0.051, P<0.01), but remained the low level after treated with PDGF alone (group P)or PDGF in combination with IL-10 (group C). Contrast to the control group, after treated with PDGF and IL-10, either alone or in combination, Bcl-2 mRNA expression was downregulated and Bax mRNA expression was up-regulated, both following the turn from group P, group I to group C.Expression of Bcl-2 mRNA in group C was significantly lower than that in group P (0.126±0.008 vs0.210±0.024, P<0.01).But no significant difference was found between group C and group I, as well as between group I and group P (P>0.05).Similarly, the expression of Bax in group C was higher than that in group P (0.513±0.016 vs0.400±0.022, P<0.01).No significant difference was found between group I and group P (P>0.05). But compared with group C, Bax expressions in group I tended to decrease (0.449±0.028 vs 0.513±0.016,P<0.05).CONCLUSION: PDGF may promote proliferation of HSCs but is neutral with respect to HSC apoptosis. IL-10 may promote the apoptosis of HSCs by up-regulating the expressions of FasL and Bax and down-regulating the expression of Bcl-2, which may be involved in its antifibrosis mechanism.展开更多
AIM: To determine the prevalence of nonalcoholic fatty liver in a specific population in Shanghai by an epidemiological survey, and to analyze risk factors of fatty liver.METHODS: Total 4009 administrative officers wh...AIM: To determine the prevalence of nonalcoholic fatty liver in a specific population in Shanghai by an epidemiological survey, and to analyze risk factors of fatty liver.METHODS: Total 4009 administrative officers who denied regular alcohol drinking participated in the survey, and underwent physical examination and laboratory tests. The important parameters were body mass index (BMI), waist hip circumferences ratio (WHR) and levels of serum lipids.Diagnosis of fatty liver was based on established real-time ultrasonographic criteria, the presence of an ultrasonographic pattern consistent with 'bright liver', with evident ultrasonographic contrast between hepatic and renal parenchyma, vessel blurring, and narrowing of the lumen of the hepatic veins. Analysis of data was performed through SPSS for Windows statistical package.RESULTS: The overall prevalence of fatty liver was 12.9 %,15.8 % in males and 7.5 % in females, and the prevalence of fatty liver in males younger than 50 years old, was significantly higher (13.3 %) than that of in females (2.7 %).But the difference between the sexes became less significant in people older than 50 years (19.1% vs 18.1%). The prevalence of fatty liver was increased with age; this was markedly presented in females younger than 50 years.Multiple variant regression analysis demonstrated that the prevalence of fatty liver was positively correlated to several risk factors, including male, aging (＞50yr), hyperlipidemia,impaired glucose tolerance/diabetes mellitus, hypertension and overweight/obesity.CONCLUSION: There is a high prevalence of nonalcoholic fatty liver among certain population in Shanghai, to which overweight and hyperlipidemia are closely relevant.展开更多
AIM: Gastroesophageal reflux disease (GERD) is a common disorder in the Western population, but detailed population-based data in China are limited. The aim of this study was to understand the epidemiology of symptoma...AIM: Gastroesophageal reflux disease (GERD) is a common disorder in the Western population, but detailed population-based data in China are limited. The aim of this study was to understand the epidemiology of symptomatic gastroesophageal reflux (SGER) in adults of Xi'an, a northwestern city of China, and to explore the potential risk factors of GERD. METHODS: Symptoms suggestive of GERD, functional dyspepsia (FD), irritable bowel syndrome (IBS), upper respiratory diseases and some potential risk factors were investigated in a face-to-face manner in a region-stratified random samples of 2 789 residents aged 18-70 years in Xi'an by using a standardized questionnaire. METHODS: With a response rate of 91.8%, the prevalence of SGER was 16.98% (95% CI, 14.2-18.92) in Xi'an adults, and no gender-related difference was observed (P<0.05). SGER was more common among subjects aged 30-70 years than in those aged 18-29 years (P<0.01). The prevalence of SGER in rural, urban and suburban subjects was 21.07%, 17.44% and 12.12%, respectively, and there was a significant difference between rural, urban and suburban regions (P<0.05). Compared with subjects without SGER, the prevalence of symptoms suggestive of FD and IBS, pneumonia, asthma, bronchitis, laryngitis, pharyngitis, chronic cough, wheeze, globus sensation, oral ulcer and snore was significantly increased in subjects with SGER (P<0.01). Heavy smoking (OR=5.76; CI, 3.70-6.67), heavy alcohol use (OR=2.85; CI, 1.67-4.49), peplJc ulcer (OR=5.76; CI, 3.99-8.32), cerebral palsy (OR=3.97; CI, 1.97-8.00), abdominal operation (OR=2.69; CI, 1.75-4.13), obesity(OR=2.16; CI, 1.47-3.16), excessive food intake (OR= 1.43;CI, 1.17-1.15), sweet food (OR=1.23; CI, 0.89-1.54), and consumption of coffee (OR= 1.23; CI, 0.17-2.00) were independently associated with SGER. The episodes of GERD were commonly precipitated by dietary factors (66.05%), followed by body posture (26.54%), ill temper (23.72%), fatigue (22.32%) and stress (10.93%). CONCLUSION: GERD is common in Xi'an's adult population with a mild or moderate degree. The etiology and pathogenesis of GERD are probably associated with FD, IBS, and some respiratory, laryngopharyngeal and odontostological diseases or symptoms. Some lifestyles, diseases and dietary factors are the risk factors of GERD.展开更多
AIM: To investigate the effects of rhubarb on severe acute pancreatitis (SAP) in rats. METHODS: Severe acute pancreatitis was induced by two intraperitoneal injections of cerulein (40 μg/kg body weight) plus 5-h rest...AIM: To investigate the effects of rhubarb on severe acute pancreatitis (SAP) in rats. METHODS: Severe acute pancreatitis was induced by two intraperitoneal injections of cerulein (40 μg/kg body weight) plus 5-h restraint water-immersion stress. Rhubarb (75-150 mg/kg) was orally fed before the first cerulein injection. The degree of pancreatic edema, serum amylase level, local pancreatic blood flow (PBF), and histological alterations were investigated. The effects of rhubarb on pancreatic exocrine secretion in this model were evaluated by comparing with those of somatostatin. RESULTS: In the Cerulein+Stress group, severe edema and diffuse hemorrhage in the pancreas were observed, the pancreatic wet weight (11.60±0.61 g/Kg) and serum amylase (458 490±43 100 U/L) were markedly increased (P<0.01 vs control). In the rhubarb (150 mg/kg) treated rats, necrosis and polymorphonuclear neutrophil (PMN) infiltration in the pancreas were significantly reduced (P<0.01), and a marked decrease (50%) in serum amylase levels was also observed (P<0.01). PBF dropped to 38% (93±5 mL/min per 100 g) of the control in the Cerulein+Stress group and partly recovered in the Cerulein+Stress+Rhubarb 150 mg group (135±12 mL/min per 100 g) (P<0.01). The pancreatic exocrine function was impaired in the SAP rats. The amylase levels of pancreatic juice were reduced in the rats treated with rhubarb or somatostatin, comparing with that of untreated SAP group. The bicarbonate concentration of pancreatic juice was markedly elevated only in the rhubarbtreated group (P<0.01). CONCLUSION: Rhubarb can exert protective effects on SAP, probably by inhibiting the inflammation of pancreas, improving pancreatic microcirculation, and altering exocrine secretion.展开更多
AIM:To detect the micrometastasis of gastric carcinoma in peripheral blood circulation using immunomagnetic beads sorting technique and RT-PCR technique,and to discuss its significance and the difference between the t...AIM:To detect the micrometastasis of gastric carcinoma in peripheral blood circulation using immunomagnetic beads sorting technique and RT-PCR technique,and to discuss its significance and the difference between the two methods.METHODS:Density gradient centrifugation was used to isolate mononuclear cells from peripheral blood,immunomagnetic beads sorting technique and RT-PCR technique were used to detect the disseminated carcinoma cells.HE,immunocytochemical and immunofluorescence staining were also used to identify the characteristics of the cells separated with immunomagnetic beads sorting technique.RESULTS:Cells expressing cytokeratin were separated and enriched from the peripheral blood specimens of patients suffering from gastric carcinoma or chronic gastritis. After HE staining, two kinds of ceils with little cytoplasm were found.Majority of these cells had small and round nuclei, even chromatins and the thickness of nuclear membrane was normal. Immunohistochemical staining indicated that there were CD34 and CD45 expression on the cell membrane ofthis kind of cells and these cells also showed expressed human telomerase reverse transcriptase by immunofluorescence staining, but the expression of carcinoembryonic antigen was absent.So,these cells might hematopoiesis precursors.Another kind of cells had larger and abnormal nuclei with thicker nuclear membranes. Massed chromatins and polynudeoli were found in the nudei. These cells expressed human telomerase reverse transcriptase and carcinoembryonic antigen,but CD34 and CD45 were not found on the cell membrane.So,these cells were considered as gastric carcinoma cells escaping from the original focuses and existing in the peripheral blood circulation.Cardnoma cells were found in 25 of 60(41.7%) specimens of peripheral blood from patients with gastric carcinoma, while there were no such cells separated from the blood specimens of chronic gastritis patients.The difference of positive rates of disseminated carcinoma cells between two groups was markedly significant (P<0.005).The expressions of CK20 mRNA in peripheral blood specimens were examinated withRT-PCR. CK20 mRNA was detected from 32 of 60(53.3%) peripheral blood specimens in the group of gastric carcinoma patients,while none of the specimens from patients suffering from chronic gastritis had CK20 mRNA. Significant difference was also found between two groups (P<0.005).Statistic analyses also showed that there was a significant difference between the positive rates of two methods in detecting the disseminated carcinoma cells from the peripheral blood circulation of gastric carcinoma patients (P<0.05).CONCLUSION:The results demonstrated that there were disseminated carcinoma cells in the peripheral blood circulation of some patients with gastric carcinoma.Disseminated carcinoma cells can be detected from the peripheral blood samples with immunomagnetic beads sorting technique and RT-PCR technique.The positive rate of RT-PCR technique is higher than that of immunomagnetic beads sorting technique in detecting micrometastasis.展开更多
AIM: To investigate the expression of matrix metallopr-oteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic fibrosis and the antifibrogenic role of exogenous interleukin-10 (IL-10). METHODS: Hepatic fibro...AIM: To investigate the expression of matrix metallopr-oteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic fibrosis and the antifibrogenic role of exogenous interleukin-10 (IL-10). METHODS: Hepatic fibrosis was induced by CCI4 administration and 60 male Sprague-Dawley rats were randomly divided into normal control group (group N, 8 rats), CCI4-induced group (group C, 28 rats) and IL-10-treated group (group I, 24 rats). At the beginning of the 7th and 11th wk, rats in each group were routinely perfused with pronase E and type IV collagenase through portal vein catheter and the suspension was centrifuged by 11% Nycodenz density gradient to isolate hepatic stellate cells (HSCs). RT-PCR was used to analyze mRNA of MMP-2 and TIMP-1 from freshly isolated cells. Densitometric data were standardized with β-actin signals. Immunocytochemistry was performed to detect MMP-2 and TIMP-1 expression in HSC cultured for 72 h. RESULTS: Compared to group N in the 7th wk, MMP-2 and TIMP-1 mRNA increased in group C (P= 0.001/0.001) and group I (P= 0.001/0.009). The level of MMP-2 and TIMP-1 mRNA in group I was significantly lower than that in group C (P= 0.001/0.001). In the 11th wk, MMP-2 mRNA in group I was still lower than that in group C (P = 0.005), but both dropped compared with that in the 7th week (P = 0.001/0.004). TIMP-1 mRNA in group I was still lower than that in group C (P= 0.001), and increased in group C (P= 0.001) while decreased in group I (P = 0.042) compared with that in the 7th wk. Same results were found by immunocytochemistry. CONCLUSION: Expression of MMP-2 and TIMP-1 is increased in hepatic fibrosis. IL-10 exhibits an antifibrogenic effect by suppressing MMP-2 and TIMP-1 expression.展开更多
AIM: To investigate the effects of IH764-3 on HSC apoptosis,and the expression of caspase-3 protein in HSC apoptoticprocess.METHODS: HSCs were cultured in medium with differentIH764-3 doses ( 10 μg@ mL-1 , 20 μg@ mL...AIM: To investigate the effects of IH764-3 on HSC apoptosis,and the expression of caspase-3 protein in HSC apoptoticprocess.METHODS: HSCs were cultured in medium with differentIH764-3 doses ( 10 μg@ mL-1 , 20 μg@ mL-1 , 30 μg@ mL-1,40μg @mL-1) and without IH764-3, and HSC proliferation wasquantitatively measured by 3 H-thymidine incorporation. Themorphological changes of HSCs were observed withtransmission electron microscope after exposure to the doseof 40 μg@ mL-1 of IH764-3 for 48 hr, The apoptosis rates weredetected by annexin V/PI and TdT-mediated dUTP nick endlabeling (TUNEL). The expression of caspase-3 protein wasdetermined by flow cytometry.RESULTS: (1) HSC proliferation rates induced with differentIH764-3 doses ( 10 μg@ mL-1 , 20 μg@ mL-1 , 30 μg@ mL-1 , 40 μg@mL-1) were significantly reduced compared with that of thecontrol group ( P＜ 0.01). (2)With the doses above, IH764-3dose-dependently produced HSC apoptosis rates of 6.7 %(9.4%),9.3 %(21.6 %),15.1%(27.2 %) and 19.0 %(28.4 %)respectively, by annexin V and PI-labeled flow cytometry assay(or TUlE L), while it was only 2.3 %(6.7 %) in the control. (3)The expression of caspase-3 protein in IH764-3 groups wassignificantly higher than that of the cortrol (P＜0.05).CONCLUSION: Within the dose range used in present study,IH764-3 can inhibit HSC proliferation, as well as enhance HSCapoptosis. Furthermore, IH764-3 can significantly increasethe caspase-3 protein expression.展开更多
AIM: To evaluate the value of miniprobe sonography (MPS),spiral CT and MR imaging (MRI) in the tumor and regionallymph nocle staging of esophageal cancer.METHODS: Eight-six patients (56 men and 30 women; agerange of 3...AIM: To evaluate the value of miniprobe sonography (MPS),spiral CT and MR imaging (MRI) in the tumor and regionallymph nocle staging of esophageal cancer.METHODS: Eight-six patients (56 men and 30 women; agerange of 39-73 years, mean 62 years) with esophagealcarcinoma were staged .preoperatively with imagingmodalities. Of them, 81 (94 %) had squamous cell carcinoma,4(5 %) adenocarcinoma, and 1(1%) adenoacanthoma.Eleven patients (12 %) had malignancy of the upper onethird, 41 (48 %) of the mid-esophagus and 34 (40 %) ofthe distal one third. Forty-one were examined by spiral CTin whom 13 were co-examined by MPS, and forty-five byMRI in whom 18 were also co-examined by MPS. Theseimaging results were compared with the findings of thehistopathologic examination for resected specimens.RESULTS: In staging the depth of tumor growth, MPS wassignificantly more accurate (84 %) than spiral CT and MRI(68 % and 60 %, respectively, P＜0.05). The specificity andsensitivity were 82 % and 85 % for MPS; 60 % and 69 % forspiral CT; and 40 % and 63 % for MRI, respectively. In stagingregional lymph nodes, spiral CT was more accurate (78 %)than MPS and MRI (71% and 64 %, respectively), but thedifference was not statistically significant. The specificity andsensitivity were 79 % and 77 % for spiral CT; 75 % and 68 %for MPS; and 68 % and 62 % for MRI, respectively.CONCLUSION:MPS is superior to spiral CT or MRI for Tstaging, especially in early esophageal cancer. However,the three modalities have the similar accuracy in N staging.Spiral CT or MRI is helpful for the detection of far-distancemetastasis in esophageal cancer.展开更多
AIM: To seek the X associated protein (XAP) with theconstructed bait vector pAS2-1X from normal human livercDNA library.METHODS: The X region of the HBV gene was amplied byPCR and cloned into the eukaryotic expression...AIM: To seek the X associated protein (XAP) with theconstructed bait vector pAS2-1X from normal human livercDNA library.METHODS: The X region of the HBV gene was amplied byPCR and cloned into the eukaryotic expression vector pAS2-l.The reconstituted plasmid pAS2-1X was transformed intothe yeast cells and the expression of X protein (pX) wasconfirmed by Western blot analysis. Yeast cells werecotransformed with pAS2-1X and the normal human livercDNA library and were grown in selective SC/-trp-leu-his-ade medium, the second screen was performed with theLacZ report gene. Furthermore, segregation analysis andmating experiment were performed to eliminate the falsepositive and the true positive clones were selected for PCRand sequencing.RESULTS: Reconstituted plasmid pAS2-1X including theanticipated fragment of X gene was proved by auto-sequencing assay. Western blot analysis showed thatreconstituted plasmid pAS2-1X expressed BD: X fusionprotein in yeast cells. Of 5 × 106 transformed coloniesscreened, 65 grew in the selective SC/-trp-leu-his-ademedium, 5 scored positive for β-gal activity, and only 2remaining clones passed through the segregation analysisand mating experiment. Sequence analysis identified thattwo clones contained similar cDNA fragment: GAACFFGCG.CONCLUSION: The short peptide (glutacid-leucine-alanine)is a possible required site for XAP binding to pX. Normalhuman liver cDNA library has difficulties in expressing theintegrated XAP on yeast cells.展开更多
AIM:There is still no accepted conclusion regarding the clinical features and related risk factors of patients with fatty liver.The large-scale clinical studies have not carried out yet in Guangzhou area. The aim of t...AIM:There is still no accepted conclusion regarding the clinical features and related risk factors of patients with fatty liver.The large-scale clinical studies have not carried out yet in Guangzhou area. The aim of the present study was to investigate the clinical features and related risk factors of patients with fatty liver in Guangzhou area.METHODS:A total of 413 cases with fatty liver were enrolled in the study from January 1998 to May 2002. Retrospective case-control study was used to evaluate the clinical features and related risk factors of fatty liver with logistic regression.RESULTS: Obesity (OR：21.204), alcohol abuse (OR：18.601),type 2 diabetes mellitus (OR：4.461), serum triglyceride (TG)(OR：3.916), serum low-density lipoprotein cholesterol (LDL-C) (OR：1.840) and fasting plasma glucose (FPG) (OR：1.535) were positively correlated to the formation of the fatty liver.The levels of serum alanine aminotransferase (ALT) and gammaglutamyltransferase (GGT) increased mildly in the patients with fatty liver and were often less than 2-fold of the normal limit.The higher abnormalities of aspartate aminotransferase (AST) levels (42.9%) with AST/ALT more than 2(17.9%) were found in patients with alcoholic fatty liver (AFL) than those with nonalcoholic fatty liver (NAFL) (16.9% and 5.0% respectively).The elevation of serum TG, cholesterol (CHOL), LDL-C was more common in patients with NAFL than with AFL.CONCLUSION:Obesity, alcohol abuse, type 2 diabetes mellitus and hyperlipidernia may be independent risk factors of fatty liver.The mildly abnormal hepatic functions can be found in patients with fatty liver.More obvious damages of liver function with AST/ALT usually more than 2 were noted in patients with AFL.展开更多
AIM:Osteopontin (OPN) is a phosphorylated glycoprotein with diverse functions including cancer development,progression and metastasis. It is unclear how osteopontin is regulated in HepG2 cells.The aim of this study wa...AIM:Osteopontin (OPN) is a phosphorylated glycoprotein with diverse functions including cancer development,progression and metastasis. It is unclear how osteopontin is regulated in HepG2 cells.The aim of this study was to investigate the effect of epidermal growth factor on the expression of osteopontin in HepG2 cells, and to explore the signal transduction pathway mediated this expression.METHODS: Osteopontin expression was detected by RNAase protection assay and Western blot. Wortmannin, a specific inhibitor of PI3K, was used to see if PI3K signal transduction was involved in the induction of osteopontin gene expression.RESULTS:HepG2 cells constitutively expressed low levels of osteopontin.Treatment with epidermal growth factor increased osteopontin mRNA and protein level in a doseand time-dependent manner.Application of wortmannin caused a dramatic reduction of epidermal growth factorinduced osteopontin expression.CONCLUSION:Osteopontin gene expression can be induced by treatment of HepG2 cells with epidermal growth factor.Epidermal growth factor may regulate osteopontin gene expression through PI3K signaling pathway. Several potential targets in the pathway can be manipulated to block the synthesis of osteopontin and inhibit liver cancer metastasis.展开更多
AIM: Cyclooxygenase (COX)-2 is over expressed ingastrointestinal neoplasm. Helicobacter pylori ( H pylon)infection is causally linked to gastric cancer. However, theexpression of COX-2 in various stages of Hpylori-ass...AIM: Cyclooxygenase (COX)-2 is over expressed ingastrointestinal neoplasm. Helicobacter pylori ( H pylon)infection is causally linked to gastric cancer. However, theexpression of COX-2 in various stages of Hpylori-associatedgastric carcinogenesis pathway has not been elucidated.Therefore, the aim of this study was to clarify the role ofH pyloriinduced COX-2 expression during carcinogenesisin the stomach.METHODS: Gastric biopsies from 138 subjects [30 casesof chronic superficial gastritis (CSG), 28 cases of gastricglandular atrophy (GA), 45 cases of gastric mucosal intestinalmetaplasia (IM), 12 cases of moderate gastric epithelialdysplasia and 23 cases of gastric cancer] were enrolled.Hpyloriinfection was assessed by a rapid urease test andhistological examination (modified Giemsa staining). Theexpression of COX-1 and COX-2 in human gastric mucosawas detected by immunohistochemical staining.I^7~I_~: Hpyloriinfeddon rate was 64.3% in GA and 69.5%in gastric cancer, which was significantly higher than that(36.7%) in CSG (P<0.05). The positive expression rates ofCOX-2 were 10.0%, 35.7%, 37.8%, 41.7% and 69.5% inCSG, GA, IM, dysplasia and gastric cancer, respectively.From CSG to GA, IM, dysplasia and finally to gastric cancer,expression of COX-2 showed an ascending tendency, whereasCOX-1 expression did not change significantly in the gastricmucosa. The level of COX-2 expression in IM and dysplasiawas significantly higher in H pylon:positive than in H pylor/-negative subjects (P<0.01).展开更多
AIM: To evaluate the expression of cyclooxygenase (COX2) and the relationship with tumor angiogenesis and advancement in gastric adenocarcinoma.METHODS: Immunohistochemical stain was used for detecting the expression ...AIM: To evaluate the expression of cyclooxygenase (COX2) and the relationship with tumor angiogenesis and advancement in gastric adenocarcinoma.METHODS: Immunohistochemical stain was used for detecting the expression of COX-2 in 45 resected specimens of gastric adenocarcinoma; the monoclonal antibody against CD34 was used for displaying vascular endothelial cells, and microvascular density (MVD) was detected by counting of CD34-positive vascular endothelial cells. Paracancerous tissues were examined as control.RESULTS: Immunohistological staining with COX-2-specific polyclonal antibody showed cytoplasmic staining in the cancer cells, some atypical hyperplasia and intestinal metaplasia,as well as angiogenic vasculature present within the tumors and prexisting vasculature adjacent to cancer lesions. The rate of expression of COX-2 and MVD index in gastric cancers were significantly increased, compared with those in the paracancerous tissues (77.78 vs 33.33 %, 58.13±19.99 vs 24.02±10.28, P＜0.01, P＜0.05, respectively). In 36 gastric carcinoma specimens with lymph node metastasis, the rate of COX-2 expression and MVD were higher than those in the specimens without metostasis (86.11 vs 44.44 %,58.60±18.24 vs 43.54±15.05, P＜0.05, P＜0.05, respectively).The rate of COX-2 expression and MVD in the specimens with invasive serosa were significantly higher than those in the specimens without invasion to serosa (87.88 vs 50.0 %,57.01±18.79 vs42.35±14.65, P＜0.05, P＜0.05). Moreover,MVD in COX-2-positive specimens was higher than that in COX-2-negative specimens (61.29±14.31 vs 45.38±12.42,P＜0.05). COX-2 expression was positively correlated with MVD (r=0.63, P＜0.05).CONCLUSION: COX-2 expression might correlate with the occurance and advancement of gastric carcinoma and is involved in tumor angiogenesis in gastric carcinoma. It is likely that COX-2 by inducing angiogenesis can be one of mechanisms which promotes invasion and metastasis of gastric carcinoma. It may become a new therapeutic target for anti-angiogenesis.展开更多
AIM：To analyze the genetic and epigenetic alterations of RUNX3 gene, a potential putative tumor suppressor gene,in hepatocellular carcinoma (HCC).METHODS: PCR-based loss of heterozygosity (LOH) detection, analysis of...AIM：To analyze the genetic and epigenetic alterations of RUNX3 gene, a potential putative tumor suppressor gene,in hepatocellular carcinoma (HCC).METHODS: PCR-based loss of heterozygosity (LOH) detection, analysis of mutation with PCR-single strand conformational polymorphism (SSCP) and sequencing, and methylation study with methylation specific PCR (MSP) were performed on RUNX3 gene in a series of 62 HCCs along with their matched normal tissues.RESULTS:Mutation of RUNX3 gene was not found, but one single nucleotide polymorphism with T to A transversion at the second nucleotide of the 18th condon was found.Nine of 26 informative cases (34.6%) showed allelic loss on the polymorphic site and 30 cases (48.4%) revealed hypermethylation of RUNX3 gene in promoter CpG islands.Furthermore,of the 9 cases with LOH, 8 (88.9%) also had hypermethylation.CONCLUSION:Our findings indicate that inactivation of RUNX3 gene through allelic loss and promoter hypermethylation might be one of the major mechanisms in hepatocellualr carcinogenesis.展开更多
AIM: To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake, DNA synthesis,collagen synthesis and secretion, inward whole-cell calcium concentration ([Ca^2+]i) as well as the blocking...AIM: To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake, DNA synthesis,collagen synthesis and secretion, inward whole-cell calcium concentration ([Ca^2+]i) as well as the blocking effect of verapamil on ET-l-stimulated release of inward calcium(Ca2+) of HSC in vitro.METHODS: Rat hepatic stellate cells (HSCs) were isolated and cultivated. ^3H-TdR and ^3H-proline incorporation used for testing DNA uptake and synthesis, collagen synthesis and secretion of HSCs cultured in vitro;, Fluorescent calcium indicator Fura-2/AM was used to measure [Ca^2+]i inwardHSCs.RESULTS: ET-1 at the concentration of 5×10^-8 mol/L,caused significant increase both in HSC DNA synthesis (2 247+344 cpm, P<0.05) and DNA uptake (P<0.05) when compared with the control group. ET-1 could also increase collagen synthesis (P<0.05 vs control group) and collagen secretion (P<0.05 vscontrol group). Besides, inward HSC [Ca^2+]i reached a peak concentration (422±98 mol/L, P<0.001)at 2 min and then went down slowly to165+51 mol/L(P<0.01) at 25 min from resting state (39±4 mol/L)after treated with ET-1. Verapamil (5 mol/L) blocked ET-1-activated [Ca^2+li inward HSCs compared with control group(P<0.05). Fura-2/AM loaded HSC was suspended in no Ca^2+ buffer containing 1 mol/L EGTA, 5 rain later, 10^-8 mol/L of ET-1 was added, [Ca^2+]i inward HSCs rose from resting state to peak 399±123 mol/L, then began to come down by the time of 20 min. It could also raise [Ca^2+]i inward HSCs even without Ca^2+ in extracellular fluid, and had a remarkable dose-effect relationship(P<0.05). Meanwhile,verapamil could restrain the action of ET-1(P<0.05).CONCLUSION: Actions of ET-1 on collagen metabolism of HSCs may depend on the transportation of inward wholecell calcium.展开更多
AIM: It has been noticed that gastroenteritis or dysentery plays a role in pathogenesis of irritable bowel syndrome (IBS), and antibiotics can increase functional abdominal symptoms, both of which may be partly due to...AIM: It has been noticed that gastroenteritis or dysentery plays a role in pathogenesis of irritable bowel syndrome (IBS), and antibiotics can increase functional abdominal symptoms, both of which may be partly due to intestinal flora disorders. This study was to determine the change of gut flora of IBS, a cluster of abdominal symptoms. Because of the chronic course and frequent occurrence of the disease, IBS patients suffered much from it. So the quality of life (Qol) of IBS patients was also evaluated in this study. METHODS: Twenty-five Rome II criteria-positive IBS patients were recruited, and 25 age and gender-matched healthy volunteers were accepted as control. The fecal flora, including Lactobacillus, Bifidobacterium, Bacteroides, C. perfringens Enterobacteriacea and Enterococus, were analyzed quantitatively and qualitatively. We also calculated the ratio of BitZdobacterium to Enterobacteriaceae ( B/E ratio) in both IBS patients and controls. In both groups, the data were further analyzed based on age difference, and comparisons were made between the younger and elder subgroups. We also evaluated the quality of life (QoL) of IBS patients and the control group using the Chinese version of SF-36 health questionnaire. RESULTS: In IBS patients, the number of fecal Bifidobacterium was significantly decreased and that of Enterobacteriaceae was significantly increased compared with that in healthy controls (both P<0.05).The mean microbial colonization resistance (CR) of the bowel in IBS patients was smaller than 1, making a significant difference compared with that in control which was more than 1 (P<0.01). There was no significant difference in gut flora between two subgroups. While in control, the elder subgroup presented more Enterobactenacea than the younger one (P<0.05). Compared with the control group, IBS patients had significantly lower scores on all SF-36 scales, with the exception of physical functioning. However, there was no significant correlation between quality of life and enteric symptoms in IBS patients. CONCLUSION: There are intestinal flora disorders in IBS patients, which may be involved in triggering the IBS-like symptoms. IBS patients experience significant impairment in QoL, however, the impairment is not caused directly by enteric symptoms.展开更多
AIM: To investigate the relationship between gastric dysmotility,gastrointestinal hormone abnormalities, and neuroendocrine cells in gastrointestinal mucosa in patients with functional dyspepsia (FD).METHODS: Gastric ...AIM: To investigate the relationship between gastric dysmotility,gastrointestinal hormone abnormalities, and neuroendocrine cells in gastrointestinal mucosa in patients with functional dyspepsia (FD).METHODS: Gastric emptying was assessed with solid radiopaque markers in 54 FD patients, and the patients were divided into two groups according to the results, one with delayed gastric emptying and the other with normal gastric emptying. Seventeen healthy volunteers acted as normal controls. Fasting and postprandial plasma levels and gastroduodenal mucosal levels of gastrointestinal hormones gastrin, somatostatin (SS) and neurotensin (NT)were measured by radioimmunoassay in all the subjects.G cells (gastrin-producing cells) and D cells (SS-producing cells) in gastric antral mucosa were immunostained with rabbit anti-gastrin polyclonal antibody and rabbit anti-SS polyclonal antibody, respectively, and analyzed quantitatively by computerized image analysis.RESULTS: The postprandial plasma gastrin levels, the fasting and postprandial plasma levels and the gastric and duodenal mucosal levels of NT were significantly higher in the FD patients with delayed gastric emptying than in those with normal gastric emptying and normal controls. The number and gray value of G and D cells and the G cell/D cell number ratio did not differ significantly between normal controls and the FD patients with or without delayed gastric emptying.CONCLUSION: Our findings suggest that the abnormalities of gastrin and NT may play a role in the pathophysiology of gastric dysmotility in FD patients, and the abnormality of postprandial plasma gastrin levels in FD patients with delayed gastric emptying is not related to the changes both in the number and gray value of G cells and in the G cell/D cell number ratio in gastric antral mucosa.展开更多
基金Supported by the Scientific Research Foundation of Chinese PLA,during the 10th-Five-Year Plan Period,No.01MA172
文摘AIM: To study the effect of NF-κB, survivin, Bd-2 and Caspase3 on tumor necrosis factors related apoptosis inducing ligand (TRAIL) induced apoptosis of gastric cancer cells. METHODS: Gastric cancer cells of SGC-7901, MKN28, MKN45 and AGS lines were cultured in PRMI-1640 medium and the apoptosis rates of the cells of 4 lines were observed after treatment of tumor necrosis factors related apoptosis indudng ligand (TRAIL) with a flow cytometer. The expression of NF-κB, survivin, Bcl-2 and Caspase3 in gastric cancer cells of 4 lines was analyzed with Western blot. RESULTS: After the gastric cancer cells were exposed to TRAIL 300 ng/ml for 24 hours, the apoptosis rate was 36.05%, 20.27%, 16.50% and 11.80% in MKN28, MKN45,AGS and SC-C-7901cells respectively. Western blot revealed that the expressions of NF-EB and survivin were lower in MKN28 cells than in MKN45, AGS and SGC-7901 cells. In contrast, the expression of Caspase3 was higher in MKN28 cells than in MKN45, AGS and SGC-7901 cells. CONCLUSION: There is a selectivity of TRAIL potency to induce apoptosis in gastric cancer cells of different cell lines.The anticancer potency of TRAIL is associated with the decreased expression of NF-κB and survivin and increased expression of Caspase3 of gastric cancer cells.
文摘AIM: To test the hypothesis that introduction of antisense TβR Ⅰ and TβR Ⅱ eukaryotic expressing plasmids into a rat model of immunologically induced liver fibrosis might block the action of TGF-β1 and halt the progression of liver fibrosis. METHODS: RT-Nest-PCR and gene recombination techniques were used to construct rat antisense TβR Ⅰ and TβR Ⅱ recombinant plasmids which could be expressed in eukaryotic cells. The recombinant plasmids and empty vector (pcDNA3) were encapsulated by glycosyl-poly-L-lysine and then transducted into rats of pig serum-induced liver fibrosis model. Expression of exogenously transfected gene was assessed by Northern blot, and hepatic expressions of TβR Ⅰ and TβR Ⅱ were evaluated by RT-PCR and Western blot.We also performed ELISA for serum TGF-β1, hydroxyproline of hepatic tissues, immunohistochemistry for collagen types Ⅰ and Ⅲ, and VG staining for pathological study of the liver tissues. RESULTS: The exogenous antisense TβR Ⅰ and TβR Ⅱ plasmids could be well expressed in vivo, and block mRNA and protein expression of TβR Ⅰ and TβR Ⅱ in the fibrotic liver at the level of mRNA respectively. These exogenous plasmid expressions reduced the level of TGF-β1 (antisense TβR Ⅰ group 23.998+3.045 ng/mL, antisense TβR Ⅱ group 23.156+3.131 ng/mL, disease control group 32.960+3.789 ng/mL; F-=-38.19, 36.73, P<0.01). Compared with disease control group, the contents of hepatic hydroxyproline (antisense TβR Ⅰ group 0.169+0.015 mg/g liver, antisense TβR Ⅱ group 0.167+0.009 mg/g liver, disease control group 0.296+0.026 mg/g liver; F=14.39, 15.48, P<0.01) and the deposition of collagen types Ⅰ and Ⅲ decreased in the two antisense treatment groups(antisense TβR Ⅰ group, collagen type Ⅰ 669.90+50.67, collagen type Ⅲ 657.29+49.48; antisense TβR Ⅱ group, collagen type Ⅰ 650.26+51.51, collagen type Ⅲ 661.58+55.28; disease control group, collagen type I 1209.44+116.60, collagen type Ⅲ 1175.14+121.44; F=15.48 to 74.89, P<0.01). Their expression also improved the pathologic classification of liver fibrosis models (compared with disease control group, X^2=17.14, 17.24, P<0.01). No difference was found in the level of TGF-β1, the contents of hepatic hydroxyproline and collagen types Ⅰ and Ⅲ and pathologic grade between pcDNA3 control group and disease control group or between the two antisense treatment groups (F=0.11 to1.06, X^2=0.13 to 0.16, P>0.05). CONCLUSION: Antisense TβR Ⅰ and TβR Ⅱ recombinant plasmids have certain reverse effects on liver fibrosis and can be used as possible candidates for gene therapy.
基金Supported by the Science and Technology Fundation of FujianProvince,No.2003D05
文摘AIM: To investigate the effects of platelet-derived growth factor(PDGF) and interleukin-10 (IL-10) on Fas/Fas-ligand and Bcl-2/Bax mRNA expressions in rat hepatic stellate cells.METHODS: Rat hepatic stellate cells (HSCs) were isolated and purified from rat liver by in situ digestion of collagenase and pronase and single-step density Nycodenz gradient.After activated by culture in vitro, HSCs were divided into 4 groups and treated with nothing (group N), PDGF (group P),IL-10 (group I) and PDGF in combinalJon with IL-10 (group C),respectively. Semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis was employed to compare the mRNA expression levels of Fas/FasL and Bcl-2/Bax in HSCs of each group.RESULTS: The expression levels of Fas between the 4 groups had no significant differences (P>0.05). FasL mRNA level in normal culture-activated HSCs (group N) was very low.It increased obviously after HSCs were treated with IL-10(group I) (0.091±0.007 vs 0.385±0.051, P<0.01), but remained the low level after treated with PDGF alone (group P)or PDGF in combination with IL-10 (group C). Contrast to the control group, after treated with PDGF and IL-10, either alone or in combination, Bcl-2 mRNA expression was downregulated and Bax mRNA expression was up-regulated, both following the turn from group P, group I to group C.Expression of Bcl-2 mRNA in group C was significantly lower than that in group P (0.126±0.008 vs0.210±0.024, P<0.01).But no significant difference was found between group C and group I, as well as between group I and group P (P>0.05).Similarly, the expression of Bax in group C was higher than that in group P (0.513±0.016 vs0.400±0.022, P<0.01).No significant difference was found between group I and group P (P>0.05). But compared with group C, Bax expressions in group I tended to decrease (0.449±0.028 vs 0.513±0.016,P<0.05).CONCLUSION: PDGF may promote proliferation of HSCs but is neutral with respect to HSC apoptosis. IL-10 may promote the apoptosis of HSCs by up-regulating the expressions of FasL and Bax and down-regulating the expression of Bcl-2, which may be involved in its antifibrosis mechanism.
文摘AIM: To determine the prevalence of nonalcoholic fatty liver in a specific population in Shanghai by an epidemiological survey, and to analyze risk factors of fatty liver.METHODS: Total 4009 administrative officers who denied regular alcohol drinking participated in the survey, and underwent physical examination and laboratory tests. The important parameters were body mass index (BMI), waist hip circumferences ratio (WHR) and levels of serum lipids.Diagnosis of fatty liver was based on established real-time ultrasonographic criteria, the presence of an ultrasonographic pattern consistent with 'bright liver', with evident ultrasonographic contrast between hepatic and renal parenchyma, vessel blurring, and narrowing of the lumen of the hepatic veins. Analysis of data was performed through SPSS for Windows statistical package.RESULTS: The overall prevalence of fatty liver was 12.9 %,15.8 % in males and 7.5 % in females, and the prevalence of fatty liver in males younger than 50 years old, was significantly higher (13.3 %) than that of in females (2.7 %).But the difference between the sexes became less significant in people older than 50 years (19.1% vs 18.1%). The prevalence of fatty liver was increased with age; this was markedly presented in females younger than 50 years.Multiple variant regression analysis demonstrated that the prevalence of fatty liver was positively correlated to several risk factors, including male, aging (＞50yr), hyperlipidemia,impaired glucose tolerance/diabetes mellitus, hypertension and overweight/obesity.CONCLUSION: There is a high prevalence of nonalcoholic fatty liver among certain population in Shanghai, to which overweight and hyperlipidemia are closely relevant.
文摘AIM: Gastroesophageal reflux disease (GERD) is a common disorder in the Western population, but detailed population-based data in China are limited. The aim of this study was to understand the epidemiology of symptomatic gastroesophageal reflux (SGER) in adults of Xi'an, a northwestern city of China, and to explore the potential risk factors of GERD. METHODS: Symptoms suggestive of GERD, functional dyspepsia (FD), irritable bowel syndrome (IBS), upper respiratory diseases and some potential risk factors were investigated in a face-to-face manner in a region-stratified random samples of 2 789 residents aged 18-70 years in Xi'an by using a standardized questionnaire. METHODS: With a response rate of 91.8%, the prevalence of SGER was 16.98% (95% CI, 14.2-18.92) in Xi'an adults, and no gender-related difference was observed (P<0.05). SGER was more common among subjects aged 30-70 years than in those aged 18-29 years (P<0.01). The prevalence of SGER in rural, urban and suburban subjects was 21.07%, 17.44% and 12.12%, respectively, and there was a significant difference between rural, urban and suburban regions (P<0.05). Compared with subjects without SGER, the prevalence of symptoms suggestive of FD and IBS, pneumonia, asthma, bronchitis, laryngitis, pharyngitis, chronic cough, wheeze, globus sensation, oral ulcer and snore was significantly increased in subjects with SGER (P<0.01). Heavy smoking (OR=5.76; CI, 3.70-6.67), heavy alcohol use (OR=2.85; CI, 1.67-4.49), peplJc ulcer (OR=5.76; CI, 3.99-8.32), cerebral palsy (OR=3.97; CI, 1.97-8.00), abdominal operation (OR=2.69; CI, 1.75-4.13), obesity(OR=2.16; CI, 1.47-3.16), excessive food intake (OR= 1.43;CI, 1.17-1.15), sweet food (OR=1.23; CI, 0.89-1.54), and consumption of coffee (OR= 1.23; CI, 0.17-2.00) were independently associated with SGER. The episodes of GERD were commonly precipitated by dietary factors (66.05%), followed by body posture (26.54%), ill temper (23.72%), fatigue (22.32%) and stress (10.93%). CONCLUSION: GERD is common in Xi'an's adult population with a mild or moderate degree. The etiology and pathogenesis of GERD are probably associated with FD, IBS, and some respiratory, laryngopharyngeal and odontostological diseases or symptoms. Some lifestyles, diseases and dietary factors are the risk factors of GERD.
文摘AIM: To investigate the effects of rhubarb on severe acute pancreatitis (SAP) in rats. METHODS: Severe acute pancreatitis was induced by two intraperitoneal injections of cerulein (40 μg/kg body weight) plus 5-h restraint water-immersion stress. Rhubarb (75-150 mg/kg) was orally fed before the first cerulein injection. The degree of pancreatic edema, serum amylase level, local pancreatic blood flow (PBF), and histological alterations were investigated. The effects of rhubarb on pancreatic exocrine secretion in this model were evaluated by comparing with those of somatostatin. RESULTS: In the Cerulein+Stress group, severe edema and diffuse hemorrhage in the pancreas were observed, the pancreatic wet weight (11.60±0.61 g/Kg) and serum amylase (458 490±43 100 U/L) were markedly increased (P<0.01 vs control). In the rhubarb (150 mg/kg) treated rats, necrosis and polymorphonuclear neutrophil (PMN) infiltration in the pancreas were significantly reduced (P<0.01), and a marked decrease (50%) in serum amylase levels was also observed (P<0.01). PBF dropped to 38% (93±5 mL/min per 100 g) of the control in the Cerulein+Stress group and partly recovered in the Cerulein+Stress+Rhubarb 150 mg group (135±12 mL/min per 100 g) (P<0.01). The pancreatic exocrine function was impaired in the SAP rats. The amylase levels of pancreatic juice were reduced in the rats treated with rhubarb or somatostatin, comparing with that of untreated SAP group. The bicarbonate concentration of pancreatic juice was markedly elevated only in the rhubarbtreated group (P<0.01). CONCLUSION: Rhubarb can exert protective effects on SAP, probably by inhibiting the inflammation of pancreas, improving pancreatic microcirculation, and altering exocrine secretion.
文摘AIM:To detect the micrometastasis of gastric carcinoma in peripheral blood circulation using immunomagnetic beads sorting technique and RT-PCR technique,and to discuss its significance and the difference between the two methods.METHODS:Density gradient centrifugation was used to isolate mononuclear cells from peripheral blood,immunomagnetic beads sorting technique and RT-PCR technique were used to detect the disseminated carcinoma cells.HE,immunocytochemical and immunofluorescence staining were also used to identify the characteristics of the cells separated with immunomagnetic beads sorting technique.RESULTS:Cells expressing cytokeratin were separated and enriched from the peripheral blood specimens of patients suffering from gastric carcinoma or chronic gastritis. After HE staining, two kinds of ceils with little cytoplasm were found.Majority of these cells had small and round nuclei, even chromatins and the thickness of nuclear membrane was normal. Immunohistochemical staining indicated that there were CD34 and CD45 expression on the cell membrane ofthis kind of cells and these cells also showed expressed human telomerase reverse transcriptase by immunofluorescence staining, but the expression of carcinoembryonic antigen was absent.So,these cells might hematopoiesis precursors.Another kind of cells had larger and abnormal nuclei with thicker nuclear membranes. Massed chromatins and polynudeoli were found in the nudei. These cells expressed human telomerase reverse transcriptase and carcinoembryonic antigen,but CD34 and CD45 were not found on the cell membrane.So,these cells were considered as gastric carcinoma cells escaping from the original focuses and existing in the peripheral blood circulation.Cardnoma cells were found in 25 of 60(41.7%) specimens of peripheral blood from patients with gastric carcinoma, while there were no such cells separated from the blood specimens of chronic gastritis patients.The difference of positive rates of disseminated carcinoma cells between two groups was markedly significant (P<0.005).The expressions of CK20 mRNA in peripheral blood specimens were examinated withRT-PCR. CK20 mRNA was detected from 32 of 60(53.3%) peripheral blood specimens in the group of gastric carcinoma patients,while none of the specimens from patients suffering from chronic gastritis had CK20 mRNA. Significant difference was also found between two groups (P<0.005).Statistic analyses also showed that there was a significant difference between the positive rates of two methods in detecting the disseminated carcinoma cells from the peripheral blood circulation of gastric carcinoma patients (P<0.05).CONCLUSION:The results demonstrated that there were disseminated carcinoma cells in the peripheral blood circulation of some patients with gastric carcinoma.Disseminated carcinoma cells can be detected from the peripheral blood samples with immunomagnetic beads sorting technique and RT-PCR technique.The positive rate of RT-PCR technique is higher than that of immunomagnetic beads sorting technique in detecting micrometastasis.
基金Supported by the Science and Technology Project of Fujian Educational Committee, No. JA04198
文摘AIM: To investigate the expression of matrix metallopr-oteinase-2 and tissue inhibitor of metalloproteinase-1 in hepatic fibrosis and the antifibrogenic role of exogenous interleukin-10 (IL-10). METHODS: Hepatic fibrosis was induced by CCI4 administration and 60 male Sprague-Dawley rats were randomly divided into normal control group (group N, 8 rats), CCI4-induced group (group C, 28 rats) and IL-10-treated group (group I, 24 rats). At the beginning of the 7th and 11th wk, rats in each group were routinely perfused with pronase E and type IV collagenase through portal vein catheter and the suspension was centrifuged by 11% Nycodenz density gradient to isolate hepatic stellate cells (HSCs). RT-PCR was used to analyze mRNA of MMP-2 and TIMP-1 from freshly isolated cells. Densitometric data were standardized with β-actin signals. Immunocytochemistry was performed to detect MMP-2 and TIMP-1 expression in HSC cultured for 72 h. RESULTS: Compared to group N in the 7th wk, MMP-2 and TIMP-1 mRNA increased in group C (P= 0.001/0.001) and group I (P= 0.001/0.009). The level of MMP-2 and TIMP-1 mRNA in group I was significantly lower than that in group C (P= 0.001/0.001). In the 11th wk, MMP-2 mRNA in group I was still lower than that in group C (P = 0.005), but both dropped compared with that in the 7th week (P = 0.001/0.004). TIMP-1 mRNA in group I was still lower than that in group C (P= 0.001), and increased in group C (P= 0.001) while decreased in group I (P = 0.042) compared with that in the 7th wk. Same results were found by immunocytochemistry. CONCLUSION: Expression of MMP-2 and TIMP-1 is increased in hepatic fibrosis. IL-10 exhibits an antifibrogenic effect by suppressing MMP-2 and TIMP-1 expression.
基金Fund of the Scientific and Technical Department of Hebei Province,No.01276134
文摘AIM: To investigate the effects of IH764-3 on HSC apoptosis,and the expression of caspase-3 protein in HSC apoptoticprocess.METHODS: HSCs were cultured in medium with differentIH764-3 doses ( 10 μg@ mL-1 , 20 μg@ mL-1 , 30 μg@ mL-1,40μg @mL-1) and without IH764-3, and HSC proliferation wasquantitatively measured by 3 H-thymidine incorporation. Themorphological changes of HSCs were observed withtransmission electron microscope after exposure to the doseof 40 μg@ mL-1 of IH764-3 for 48 hr, The apoptosis rates weredetected by annexin V/PI and TdT-mediated dUTP nick endlabeling (TUNEL). The expression of caspase-3 protein wasdetermined by flow cytometry.RESULTS: (1) HSC proliferation rates induced with differentIH764-3 doses ( 10 μg@ mL-1 , 20 μg@ mL-1 , 30 μg@ mL-1 , 40 μg@mL-1) were significantly reduced compared with that of thecontrol group ( P＜ 0.01). (2)With the doses above, IH764-3dose-dependently produced HSC apoptosis rates of 6.7 %(9.4%),9.3 %(21.6 %),15.1%(27.2 %) and 19.0 %(28.4 %)respectively, by annexin V and PI-labeled flow cytometry assay(or TUlE L), while it was only 2.3 %(6.7 %) in the control. (3)The expression of caspase-3 protein in IH764-3 groups wassignificantly higher than that of the cortrol (P＜0.05).CONCLUSION: Within the dose range used in present study,IH764-3 can inhibit HSC proliferation, as well as enhance HSCapoptosis. Furthermore, IH764-3 can significantly increasethe caspase-3 protein expression.
文摘AIM: To evaluate the value of miniprobe sonography (MPS),spiral CT and MR imaging (MRI) in the tumor and regionallymph nocle staging of esophageal cancer.METHODS: Eight-six patients (56 men and 30 women; agerange of 39-73 years, mean 62 years) with esophagealcarcinoma were staged .preoperatively with imagingmodalities. Of them, 81 (94 %) had squamous cell carcinoma,4(5 %) adenocarcinoma, and 1(1%) adenoacanthoma.Eleven patients (12 %) had malignancy of the upper onethird, 41 (48 %) of the mid-esophagus and 34 (40 %) ofthe distal one third. Forty-one were examined by spiral CTin whom 13 were co-examined by MPS, and forty-five byMRI in whom 18 were also co-examined by MPS. Theseimaging results were compared with the findings of thehistopathologic examination for resected specimens.RESULTS: In staging the depth of tumor growth, MPS wassignificantly more accurate (84 %) than spiral CT and MRI(68 % and 60 %, respectively, P＜0.05). The specificity andsensitivity were 82 % and 85 % for MPS; 60 % and 69 % forspiral CT; and 40 % and 63 % for MRI, respectively. In stagingregional lymph nodes, spiral CT was more accurate (78 %)than MPS and MRI (71% and 64 %, respectively), but thedifference was not statistically significant. The specificity andsensitivity were 79 % and 77 % for spiral CT; 75 % and 68 %for MPS; and 68 % and 62 % for MRI, respectively.CONCLUSION:MPS is superior to spiral CT or MRI for Tstaging, especially in early esophageal cancer. However,the three modalities have the similar accuracy in N staging.Spiral CT or MRI is helpful for the detection of far-distancemetastasis in esophageal cancer.
文摘AIM: To seek the X associated protein (XAP) with theconstructed bait vector pAS2-1X from normal human livercDNA library.METHODS: The X region of the HBV gene was amplied byPCR and cloned into the eukaryotic expression vector pAS2-l.The reconstituted plasmid pAS2-1X was transformed intothe yeast cells and the expression of X protein (pX) wasconfirmed by Western blot analysis. Yeast cells werecotransformed with pAS2-1X and the normal human livercDNA library and were grown in selective SC/-trp-leu-his-ade medium, the second screen was performed with theLacZ report gene. Furthermore, segregation analysis andmating experiment were performed to eliminate the falsepositive and the true positive clones were selected for PCRand sequencing.RESULTS: Reconstituted plasmid pAS2-1X including theanticipated fragment of X gene was proved by auto-sequencing assay. Western blot analysis showed thatreconstituted plasmid pAS2-1X expressed BD: X fusionprotein in yeast cells. Of 5 × 106 transformed coloniesscreened, 65 grew in the selective SC/-trp-leu-his-ademedium, 5 scored positive for β-gal activity, and only 2remaining clones passed through the segregation analysisand mating experiment. Sequence analysis identified thattwo clones contained similar cDNA fragment: GAACFFGCG.CONCLUSION: The short peptide (glutacid-leucine-alanine)is a possible required site for XAP binding to pX. Normalhuman liver cDNA library has difficulties in expressing theintegrated XAP on yeast cells.
基金Supported by the National Natural Science Foundation of China,No.30270371
文摘AIM:There is still no accepted conclusion regarding the clinical features and related risk factors of patients with fatty liver.The large-scale clinical studies have not carried out yet in Guangzhou area. The aim of the present study was to investigate the clinical features and related risk factors of patients with fatty liver in Guangzhou area.METHODS:A total of 413 cases with fatty liver were enrolled in the study from January 1998 to May 2002. Retrospective case-control study was used to evaluate the clinical features and related risk factors of fatty liver with logistic regression.RESULTS: Obesity (OR：21.204), alcohol abuse (OR：18.601),type 2 diabetes mellitus (OR：4.461), serum triglyceride (TG)(OR：3.916), serum low-density lipoprotein cholesterol (LDL-C) (OR：1.840) and fasting plasma glucose (FPG) (OR：1.535) were positively correlated to the formation of the fatty liver.The levels of serum alanine aminotransferase (ALT) and gammaglutamyltransferase (GGT) increased mildly in the patients with fatty liver and were often less than 2-fold of the normal limit.The higher abnormalities of aspartate aminotransferase (AST) levels (42.9%) with AST/ALT more than 2(17.9%) were found in patients with alcoholic fatty liver (AFL) than those with nonalcoholic fatty liver (NAFL) (16.9% and 5.0% respectively).The elevation of serum TG, cholesterol (CHOL), LDL-C was more common in patients with NAFL than with AFL.CONCLUSION:Obesity, alcohol abuse, type 2 diabetes mellitus and hyperlipidernia may be independent risk factors of fatty liver.The mildly abnormal hepatic functions can be found in patients with fatty liver.More obvious damages of liver function with AST/ALT usually more than 2 were noted in patients with AFL.
文摘AIM:Osteopontin (OPN) is a phosphorylated glycoprotein with diverse functions including cancer development,progression and metastasis. It is unclear how osteopontin is regulated in HepG2 cells.The aim of this study was to investigate the effect of epidermal growth factor on the expression of osteopontin in HepG2 cells, and to explore the signal transduction pathway mediated this expression.METHODS: Osteopontin expression was detected by RNAase protection assay and Western blot. Wortmannin, a specific inhibitor of PI3K, was used to see if PI3K signal transduction was involved in the induction of osteopontin gene expression.RESULTS:HepG2 cells constitutively expressed low levels of osteopontin.Treatment with epidermal growth factor increased osteopontin mRNA and protein level in a doseand time-dependent manner.Application of wortmannin caused a dramatic reduction of epidermal growth factorinduced osteopontin expression.CONCLUSION:Osteopontin gene expression can be induced by treatment of HepG2 cells with epidermal growth factor.Epidermal growth factor may regulate osteopontin gene expression through PI3K signaling pathway. Several potential targets in the pathway can be manipulated to block the synthesis of osteopontin and inhibit liver cancer metastasis.
基金Supported by the Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry,No.9247342057
文摘AIM: Cyclooxygenase (COX)-2 is over expressed ingastrointestinal neoplasm. Helicobacter pylori ( H pylon)infection is causally linked to gastric cancer. However, theexpression of COX-2 in various stages of Hpylori-associatedgastric carcinogenesis pathway has not been elucidated.Therefore, the aim of this study was to clarify the role ofH pyloriinduced COX-2 expression during carcinogenesisin the stomach.METHODS: Gastric biopsies from 138 subjects [30 casesof chronic superficial gastritis (CSG), 28 cases of gastricglandular atrophy (GA), 45 cases of gastric mucosal intestinalmetaplasia (IM), 12 cases of moderate gastric epithelialdysplasia and 23 cases of gastric cancer] were enrolled.Hpyloriinfection was assessed by a rapid urease test andhistological examination (modified Giemsa staining). Theexpression of COX-1 and COX-2 in human gastric mucosawas detected by immunohistochemical staining.I^7~I_~: Hpyloriinfeddon rate was 64.3% in GA and 69.5%in gastric cancer, which was significantly higher than that(36.7%) in CSG (P<0.05). The positive expression rates ofCOX-2 were 10.0%, 35.7%, 37.8%, 41.7% and 69.5% inCSG, GA, IM, dysplasia and gastric cancer, respectively.From CSG to GA, IM, dysplasia and finally to gastric cancer,expression of COX-2 showed an ascending tendency, whereasCOX-1 expression did not change significantly in the gastricmucosa. The level of COX-2 expression in IM and dysplasiawas significantly higher in H pylon:positive than in H pylor/-negative subjects (P<0.01).
文摘AIM: To evaluate the expression of cyclooxygenase (COX2) and the relationship with tumor angiogenesis and advancement in gastric adenocarcinoma.METHODS: Immunohistochemical stain was used for detecting the expression of COX-2 in 45 resected specimens of gastric adenocarcinoma; the monoclonal antibody against CD34 was used for displaying vascular endothelial cells, and microvascular density (MVD) was detected by counting of CD34-positive vascular endothelial cells. Paracancerous tissues were examined as control.RESULTS: Immunohistological staining with COX-2-specific polyclonal antibody showed cytoplasmic staining in the cancer cells, some atypical hyperplasia and intestinal metaplasia,as well as angiogenic vasculature present within the tumors and prexisting vasculature adjacent to cancer lesions. The rate of expression of COX-2 and MVD index in gastric cancers were significantly increased, compared with those in the paracancerous tissues (77.78 vs 33.33 %, 58.13±19.99 vs 24.02±10.28, P＜0.01, P＜0.05, respectively). In 36 gastric carcinoma specimens with lymph node metastasis, the rate of COX-2 expression and MVD were higher than those in the specimens without metostasis (86.11 vs 44.44 %,58.60±18.24 vs 43.54±15.05, P＜0.05, P＜0.05, respectively).The rate of COX-2 expression and MVD in the specimens with invasive serosa were significantly higher than those in the specimens without invasion to serosa (87.88 vs 50.0 %,57.01±18.79 vs42.35±14.65, P＜0.05, P＜0.05). Moreover,MVD in COX-2-positive specimens was higher than that in COX-2-negative specimens (61.29±14.31 vs 45.38±12.42,P＜0.05). COX-2 expression was positively correlated with MVD (r=0.63, P＜0.05).CONCLUSION: COX-2 expression might correlate with the occurance and advancement of gastric carcinoma and is involved in tumor angiogenesis in gastric carcinoma. It is likely that COX-2 by inducing angiogenesis can be one of mechanisms which promotes invasion and metastasis of gastric carcinoma. It may become a new therapeutic target for anti-angiogenesis.
文摘AIM：To analyze the genetic and epigenetic alterations of RUNX3 gene, a potential putative tumor suppressor gene,in hepatocellular carcinoma (HCC).METHODS: PCR-based loss of heterozygosity (LOH) detection, analysis of mutation with PCR-single strand conformational polymorphism (SSCP) and sequencing, and methylation study with methylation specific PCR (MSP) were performed on RUNX3 gene in a series of 62 HCCs along with their matched normal tissues.RESULTS:Mutation of RUNX3 gene was not found, but one single nucleotide polymorphism with T to A transversion at the second nucleotide of the 18th condon was found.Nine of 26 informative cases (34.6%) showed allelic loss on the polymorphic site and 30 cases (48.4%) revealed hypermethylation of RUNX3 gene in promoter CpG islands.Furthermore,of the 9 cases with LOH, 8 (88.9%) also had hypermethylation.CONCLUSION:Our findings indicate that inactivation of RUNX3 gene through allelic loss and promoter hypermethylation might be one of the major mechanisms in hepatocellualr carcinogenesis.
基金Supported by the Grant for Nature and Science from Shanghai,No.03ZR 14097
文摘AIM: To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake, DNA synthesis,collagen synthesis and secretion, inward whole-cell calcium concentration ([Ca^2+]i) as well as the blocking effect of verapamil on ET-l-stimulated release of inward calcium(Ca2+) of HSC in vitro.METHODS: Rat hepatic stellate cells (HSCs) were isolated and cultivated. ^3H-TdR and ^3H-proline incorporation used for testing DNA uptake and synthesis, collagen synthesis and secretion of HSCs cultured in vitro;, Fluorescent calcium indicator Fura-2/AM was used to measure [Ca^2+]i inwardHSCs.RESULTS: ET-1 at the concentration of 5×10^-8 mol/L,caused significant increase both in HSC DNA synthesis (2 247+344 cpm, P<0.05) and DNA uptake (P<0.05) when compared with the control group. ET-1 could also increase collagen synthesis (P<0.05 vs control group) and collagen secretion (P<0.05 vscontrol group). Besides, inward HSC [Ca^2+]i reached a peak concentration (422±98 mol/L, P<0.001)at 2 min and then went down slowly to165+51 mol/L(P<0.01) at 25 min from resting state (39±4 mol/L)after treated with ET-1. Verapamil (5 mol/L) blocked ET-1-activated [Ca^2+li inward HSCs compared with control group(P<0.05). Fura-2/AM loaded HSC was suspended in no Ca^2+ buffer containing 1 mol/L EGTA, 5 rain later, 10^-8 mol/L of ET-1 was added, [Ca^2+]i inward HSCs rose from resting state to peak 399±123 mol/L, then began to come down by the time of 20 min. It could also raise [Ca^2+]i inward HSCs even without Ca^2+ in extracellular fluid, and had a remarkable dose-effect relationship(P<0.05). Meanwhile,verapamil could restrain the action of ET-1(P<0.05).CONCLUSION: Actions of ET-1 on collagen metabolism of HSCs may depend on the transportation of inward wholecell calcium.
基金Supported by Shanghai Sine Pharmaceutical.Co.,Ltd.and Wenzhou Science and Technology Bureau,Zhejiang Province,China
文摘AIM: It has been noticed that gastroenteritis or dysentery plays a role in pathogenesis of irritable bowel syndrome (IBS), and antibiotics can increase functional abdominal symptoms, both of which may be partly due to intestinal flora disorders. This study was to determine the change of gut flora of IBS, a cluster of abdominal symptoms. Because of the chronic course and frequent occurrence of the disease, IBS patients suffered much from it. So the quality of life (Qol) of IBS patients was also evaluated in this study. METHODS: Twenty-five Rome II criteria-positive IBS patients were recruited, and 25 age and gender-matched healthy volunteers were accepted as control. The fecal flora, including Lactobacillus, Bifidobacterium, Bacteroides, C. perfringens Enterobacteriacea and Enterococus, were analyzed quantitatively and qualitatively. We also calculated the ratio of BitZdobacterium to Enterobacteriaceae ( B/E ratio) in both IBS patients and controls. In both groups, the data were further analyzed based on age difference, and comparisons were made between the younger and elder subgroups. We also evaluated the quality of life (QoL) of IBS patients and the control group using the Chinese version of SF-36 health questionnaire. RESULTS: In IBS patients, the number of fecal Bifidobacterium was significantly decreased and that of Enterobacteriaceae was significantly increased compared with that in healthy controls (both P<0.05).The mean microbial colonization resistance (CR) of the bowel in IBS patients was smaller than 1, making a significant difference compared with that in control which was more than 1 (P<0.01). There was no significant difference in gut flora between two subgroups. While in control, the elder subgroup presented more Enterobactenacea than the younger one (P<0.05). Compared with the control group, IBS patients had significantly lower scores on all SF-36 scales, with the exception of physical functioning. However, there was no significant correlation between quality of life and enteric symptoms in IBS patients. CONCLUSION: There are intestinal flora disorders in IBS patients, which may be involved in triggering the IBS-like symptoms. IBS patients experience significant impairment in QoL, however, the impairment is not caused directly by enteric symptoms.
文摘AIM: To investigate the relationship between gastric dysmotility,gastrointestinal hormone abnormalities, and neuroendocrine cells in gastrointestinal mucosa in patients with functional dyspepsia (FD).METHODS: Gastric emptying was assessed with solid radiopaque markers in 54 FD patients, and the patients were divided into two groups according to the results, one with delayed gastric emptying and the other with normal gastric emptying. Seventeen healthy volunteers acted as normal controls. Fasting and postprandial plasma levels and gastroduodenal mucosal levels of gastrointestinal hormones gastrin, somatostatin (SS) and neurotensin (NT)were measured by radioimmunoassay in all the subjects.G cells (gastrin-producing cells) and D cells (SS-producing cells) in gastric antral mucosa were immunostained with rabbit anti-gastrin polyclonal antibody and rabbit anti-SS polyclonal antibody, respectively, and analyzed quantitatively by computerized image analysis.RESULTS: The postprandial plasma gastrin levels, the fasting and postprandial plasma levels and the gastric and duodenal mucosal levels of NT were significantly higher in the FD patients with delayed gastric emptying than in those with normal gastric emptying and normal controls. The number and gray value of G and D cells and the G cell/D cell number ratio did not differ significantly between normal controls and the FD patients with or without delayed gastric emptying.CONCLUSION: Our findings suggest that the abnormalities of gastrin and NT may play a role in the pathophysiology of gastric dysmotility in FD patients, and the abnormality of postprandial plasma gastrin levels in FD patients with delayed gastric emptying is not related to the changes both in the number and gray value of G cells and in the G cell/D cell number ratio in gastric antral mucosa.
