AIM:To study the expression of vascular endothelial growth factor C (VEGF-C) and chemokine receptor CCR7 in gastric carcinoma and to investigate their associations with lymph node metastasis of gastric carcinoma and t...AIM:To study the expression of vascular endothelial growth factor C (VEGF-C) and chemokine receptor CCR7 in gastric carcinoma and to investigate their associations with lymph node metastasis of gastric carcinoma and their values in predicting lymph node metastasis.METHODS:The expression of VEGF-C and CCR7 in gastric carcinoma tissues obtained from 118 patients who underwent curative gastrectomy was examined by immunohistochemisty.Among these patients,39 patients underwent multi-slice spiral CT (MSCT) examination.RESULTS:VEGF-C and CCR7 were positively expressed in 52.5 and 53.4% of patients. VEGF-C expression was more frequently found in tumors with lymph node metastasis than those without it (P<0.001).VEGF-C expression was also closely related to lymphatic invasion (P<0.001), vascular invasion (P<0.01),and TNM stage (P<0.001). However,there was no significant correlation between VEGF-C expression and age at surgery, gender, tumor size, tumor location,Lauren classification,and depth of invasion. CCR7 expression was significantly higher in patients with lymph node metastasis compared with those without lymph node metastasis (P<0.001) and was also associated with tumorsize (P<0.01), depth of invasion (P<0.001), lymphatic invasion (P<0.001),and TNM stage (P<0.001).However,the presence of CCR7 had no correlation to age at surgery,gender, tumor location, Lauren classification,and vascular invasion. Among the 39 patients who underwent MSCT examination,only CCR7 expression was related to lymph node metastasis determined by MSCT (P<0.05).In the current retrospective study,the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of VEGF-C and CCR7 expression in the diagnosis of lymph node metastasis for patients with gastric carcinoma were 73.8%, 70.2%,72.6%,71.4% and 72.0%,and 82.0%,77.2%,79.4%,80.0% and 79.7%,respectively.After subdivision according to the combination of VEGF-C and CCR7 expression, receiver operating characteristic (ROC) analysis showed that the accuracy of the combined examination of VEGF-C and CCR7 expression in predicting lymph node metastasis was relatively high (area under ROC curve [Az]=0.83).CONCLUSION:The expression of VEGF-C and CCR7 is related to lymph node metastasis of gastric carcinoma and both of them may become new targets for the treatment of gastric carcinoma.Furthermore,the combined examination of VEGF-C and CCR7 expression in endoscopic biopsy specimens may be useful in predicting lymph node metastasis of gastric carcinoma and deciding the extent of surgical lymph node resection.展开更多
AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease.METHODS: Gallblad...AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease.METHODS: Gallbladder motility was studied by ultrasonography in 33 patients with gallbladder stone and 10 health subjects as controls. Plasma CCK concentration was measured by radioimmunoassay in fasting status (CCK-f) and in 30 min after lipid test meal (CCK-30).Radioligand method was employed to analyze the amount and activity of CCK-R from 33 gallstone patients having cholecystectomy and 8 persons without gallstone died of severe trauma as controls.RESULTS: The percentage of cholesterol in the gallstone composition was more than 70%. The cholesterol stone type was indicated for the patients with gallbladder stone in this study. Based on the criterion of gallbladder residual fraction of the control group, 33 gallstone patients were divided into two subgroups, contractor group (14 cases)and non-contractor group (19 cases), The concentration of CCK-30 was significantly higher in non-contractor group than that in both contractor group and control group (55.86±3.86 pmol/l vs 37.85±0.88 pmol/l and 37.95±0.74 pmol/L, P＜0.01), but there was no difference between contractor group and control group. Meanwhile no significant difference of the concentration of CCK-f could be observed among three groups. The amount of CCK-R was lower in non-contractor group than those in both control group and contractor group (10.27±0.94 fmol/mg vs24.59±2.39 fmol/mg and 22.66±0.55 fmol/mg, P＜0.01).The activity of CCK-R shown as KD in non-contractor group decreased compared to that in control group and contractor group. Only was the activity of CCK-R lower in contractor group than that in control group. The ejection fraction correlated closely with the amount of CCK-R (r = 0.9683,P＜0.01), and the concentration of CCK-30 correlated negatively with the amount of CCK-R closely (r = -0.9627,P＜0.01).CONCLUSION: The distinctive interactive relationship of gallbladder emptying, plasma CCK and CCK-R in gallbladder from this study suggested that the defect of CCK-R may be a key point leading to the impairment of gallbladder motor function and the pathogenesis of cholesterol gallstoneformation may differ in two subgroups of gallstone patient,gallbladder non-contractor group or contractor group.展开更多
AIM: To investigate the clinical characteristics and prognostic factors of a consecutive series of patients with severe acute pancreatitis (SAP). METHODS: Clinical data of SAP patients admitted to our hospital from Ja...AIM: To investigate the clinical characteristics and prognostic factors of a consecutive series of patients with severe acute pancreatitis (SAP). METHODS: Clinical data of SAP patients admitted to our hospital from January 2003 to January 2004 were retrospectively reviewed. Collected data included the age, gender, etiology, length of hospitalization, APACHE Ⅱ score at admission, local and organ/systemic complications of the patients. RESULTS: Of the 268 acute pancreatitis patients, 94 developed SAP. The mean age of SAP patients was 52 years, the commonest etiology was cholelithiasis (45.7%), the mean length of hospitalization was 70 d, the mean score of APACHE Ⅱ was 7.7. Fifty-four percent of the patients developed necrosis, 25% abscess, 58% organ/systemic failure. A total of 23.4% (22/94) of the SAP patients died. Respiratory failure was the most common organ dysfunction (90.9%) in deceased SAP patients, followed by cardiovascular failure (86.4%), renal failure (50.0%). In the SAP patients, 90.9% (20/22) developed multiple organ/systemic failures. There were significant differences in age, length of hospitalization, APACHE Ⅱ score and incidences of respiratory failure, renal failure, cardiovascular failure and hematological failure between deceased SAP patients and survived SAP patients. By multivariate logistic regression analysis, independent prognostic factors for mortality were respiratory failure, cardiovascular failure and renal failure. CONCLUSION: SAP patients are characterized by advanced age, high APACHE Ⅱ score, organ failure and their death is mainly due to multiple organ/systemic failures. In patients with SAP, respiratory, cardiovascular and renal failures can predict the fatal outcome and more attention should be paid to their clinical evaluation.展开更多
AIM:To determine the expression levels of three metabolic enzymes of fluoropyrimidines:thymidylate synthase (TS),thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) in seven human gastrointestinal c...AIM:To determine the expression levels of three metabolic enzymes of fluoropyrimidines:thymidylate synthase (TS),thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) in seven human gastrointestinal cancer cell lines,and to compare the enzyme levels with the sensitivity to 5-fluorouracil (5-FU) and 5-fluoro-2′-deoxyuridine (FdUrd).METHODS:TS,TP and DPD mRNA levels were assessed by semi-quantitative RT-PCR,TP and DPD protein contents were measured by ELISA. Fifty percent inhibitory concentrations of growth (IC50),representing the sensitivity to drugs,were determined by MTT assay.RESULTS:IC50 values ranged from 1.28 to 12.26μM for 5-FU,and from 5.02 to 24.21μM for FdUrd,respectively.Cell lines with lower DPD mRNA and protein levels tended to be more sensitive to 5-FU (P<0.05), but neither TS nor TP correlated with 5-FU IC50 (P>0.05).Only TS mRNA level was sharply related with FdUrd sensitivity (P<0.05),but TP and DPD were not (P>0.05).A correlation was found between mRNA and protein levels of DPD (P<0.05),but not TP (P<0.05).CONCLUSION:DPD and TS enzyme levels may be useful indicators in predicting the antitumor activity of 5-FU or FdUrd,respectively.展开更多
AIM: To investigate the relationship between gallbladder stone disease (GSD) and single nucleotide polymorphisms of cholesterol 7α-hydroxylase (CYP7A) gene promoter,apolipoprotein (APO) B gene exon 26, APOEgene exon ...AIM: To investigate the relationship between gallbladder stone disease (GSD) and single nucleotide polymorphisms of cholesterol 7α-hydroxylase (CYP7A) gene promoter,apolipoprotein (APO) B gene exon 26, APOEgene exon 4 or microsatellite polymorphism of low density lipoprotein receptor (LDLR) gene exon 18.METHODS: Genotypes of CYP7A, APOB, APOE and LDLR genes were determined in 105 patients with GSD diagnosed by B-mode ultrasonography and 274 control subjects.Serum lipids were analyzed with HITACHI 7060 automaic biochemical analyzer.RESULTS: Body mass index (BMI) was significantly higher in patients with GSD (24.47±3.09) than in controls (23.50±2.16).Plasma total cholesterol was lower in patients with GSD (4.66±0.92 mmol/L) than in controls (4.91±0.96 mmol/L),P<0.01 after adjusted for age, sex and BMI. The significantly higher frequency of A allele of CYP7A gene polymorphism and X+ allele of APOBgene polymorphism was seen in GSD patients. Percentages of A allele in patients and controls were 62.86% and 54.38% (P <0.05) and those of X+ allele 8.57% and 4.01% (P<0.01). Subjects with A allele had significantly lower plasma total cholesterol and LDL cholesterol than subjects with CC homozygote. In a multiple variable logistic regression model, the BMI (OR=1.13, 95% CI: 1.05-1.22), A allele (OR=1.48, 95% CI: 1.05-2.09) and X+ allele (OR=2.28, 95% CI: 1.14-4.59) were positively associated with GSD (P <0.05). Plasma total cholesterol (OR=0.69, 95% CI: 0.64-0.74) was negatively related to GSD (P<0.05).CONCLUSION: With an association analysis, it was determined that A allele of CYP7A gene and X+ allele of APOB gene might be considered as risk genes for GSD. These alleles are related with differences of serum lipids among subjects.Multiple-variable logistic regression model analysis showed that besides BMI, GSD was affected by polygenetic factors.But the mechanism for these two alleles responsible for GSD requires further investigations.展开更多
AIM:To investigate the immunotherapeutic potential of vaccine consisting of dendritic cells (DCs) pulsed with total RNA from MFC gastric cancer cells.METHODS:DCs were prepared from the spleens of strain 615 mice by ma...AIM:To investigate the immunotherapeutic potential of vaccine consisting of dendritic cells (DCs) pulsed with total RNA from MFC gastric cancer cells.METHODS:DCs were prepared from the spleens of strain 615 mice by magnetic cell sorting (MACS). Alter culture for 24h, DCs were pulsed with total RNA from MFC gastric cancer cells. Mice of one group were immunized with tumor RNA pulsed DC (RNA/DC) at the dosage of 1×10^6 on d 14 and 7 by s c inoculation before tumor implantation. Mice of another group were immunized with unpulsed DC (UDC) at the same dosage on days as the RNA/DC group. The third group of control mice was untreated. On d 0, all the mice were challenged with s c injections of 5×10^5 MFC gastric cancer cells.Alter inoculation, the mice were monitored closely with respect to tumor growth. Activities of NK cells in PBL and splenocytes and CTL were tested.RESULTS:On d 21 alter tumor cell inoculation, the mice of control group manifested the largest tumors with volume at a mean of 2.6323±1.1435cm^3, followed by the UDC and RNA/DC groups with mean volumes at 0.7536±0.3659cm^3 and 0.3688±0.6571cm^3, respectively. The activities of NK cells in PBL and splenocytes in RNA/DC group were 66.2% and 65.4%, respectively, higher than that in the control group. The tumor specific CTL activity in RNA/DC group was 49.5%, higher than that in the control group.CONCLUSION:The tumor vaccine with DCs pulsed with total RNA from gastric cancer cells possesses the ability to stimulate tumor specific CTL activity and to establish antitumor immunity when administered in vivo.展开更多
AIM: Cancer gene therapy has received more and moreattentions in the recent decade. Various systems of genetherapy for cancer have been developed. One of the mostpromising choices is the suicide gene. The product ofth...AIM: Cancer gene therapy has received more and moreattentions in the recent decade. Various systems of genetherapy for cancer have been developed. One of the mostpromising choices is the suicide gene. The product ofthymidine kinase (TK) gene can convert ganciclovir (GCV)to phosphorylated GCV, which inhibits the synthesis of cellDNA, and then induces the cells to death. Cytolines play animportant role in anti-tumor immunity. This experiment wasdesigned to combine theTK gene and mIL-2/mGM-CSFgenes to treat gastric cancer, and was expected to producea marked anti-tumor effect.METHODS: TK gene was constructed into the retroviralvector pLxSN, and the mIL-2 and mGM-CSF genes wereinserted into the eukaryotic expressing vector pIRES. Thegastric cancer cells were transfected by retroviral serum thatwas harvested from the package cells. In vitro study, thetransfected gastric cancer cells were maintained in the GCV-contained medium, to assay the cell killing effect andbystander effect. In vivo experiment, retroviral serum andcytokines plasmid were transfected into tumor-bearing mice,to observe the changes of tumor volumes and survival ofthe mice.RESULTS: In vitro experiment, 20 % TK gene transducedcells could cause 70-80 % of total cells to death. In vivoresults showed that there was no treatment effect in controlgroup and TK/GCV could inhibit the tumor growth. Thestrongest anti-tumor effect was shown in TK+mIL-2+mGM-CSF group. The pathologic examination showed necrosis ofthe cancer in the treated groups.CONCLUSION: TK/GCV can kill tumor cells and inhibit thetumor growth in vivo IL-2 and GM-CSF strongly enhancethe anti-tumor effect. Through the retrovirus and liposomemethods, the suicide gene and cytokine genes are allexpressed in the tissues.展开更多
AIM: To evaluate the effect of reactive oxygen species suchas hydrogen peroxide on the progression of human coloncancer.METHODS: Human colon carcinoma cell lines, LS174T andHCT8, were treated respectively with 10- 5,1...AIM: To evaluate the effect of reactive oxygen species suchas hydrogen peroxide on the progression of human coloncancer.METHODS: Human colon carcinoma cell lines, LS174T andHCT8, were treated respectively with 10- 5,10- 7 or 10- 9 mol@L- 1 hydrogen peroxide for 24h, and co-cultured with humanendothelial cell line ECV-304. The migration of ECV-304induced by cancer cells was calculated and the expressionlevel of vascular endothelial growth factor in cancer cellswas determined by RT-PCR analysis and ELISA.Dactinomycin of 1.5mg@ L-1 which could block transcriptionof cancer cells was applied to observe the effects of H2O2 ontranscriptional activity and the relative half-life of VEGFmRNA. Finally, to evaluate the effect H2O2 on NF-κB activityin colon cancer cells, NF-κB in cytoplasm and nucleus of thecells were detected with FITC-tagged antibody and itspresence in the nucleus (Fn ) Vs cytoplasm ( Fc ) wasmonitored by measuring the green fluorescence integratedover the nucleus by laser scanning cytometry(LSC).RESULTS: Exogenouse hydrogen peroxide of lowconcentration increased the migration of endothelial cellsinduced by colon cancer cells. When cancer cells weretreated with 10-5 mol@ L-1 H2O2, the migration number ofendothelial cells induced by LS174T cells was 203 ± 70, andthe number induced by HCT8 cells was 145 ± 65. The twovalues were significantly higher than those treated with otherconcentrations of h2O2 ( P ＜ 0.01 ). The expression ofvascular endothelial growth factor in cancer cells, whichcould be blocked by dactinomycin, were increased to acertain degree, while the relative half-life of VEGF mRNAwas not prolonged after treatment with hydrogen peroxide.The activity of NF-κB in colon cells rose after the cells wereexposed to hydrogen peroxide for 24h. The Fn values inHCT8 cells were 91 ± 13 (0 mol@ L- 1 h2O2) and 149 ± 40( 10-5mol@L-1 h2O2) ( P＜ 0.05), in LS174T cells were 127 ± 35(0mol@L-1 H2O2) and 192± 11(10-5 mol@L-1 h2O2) (P＜ 0.05).lt is similar to the case of VEGF expression in cancer cells.CONCLUSION: Hydrogen peroxide increases vascularendothelial growth factor expression in colon cancer cells,and it is likely that reactive oxygen species such ashydrogen peroxide facilitates the development of coloncancer.展开更多
AIM: Laparoscopic surgery, especially laparoscopic rectal surgery, for colorectal cancer has been developed considerably. However, due to relatively complicated anatomy and high requirements for surgery techniques, la...AIM: Laparoscopic surgery, especially laparoscopic rectal surgery, for colorectal cancer has been developed considerably. However, due to relatively complicated anatomy and high requirements for surgery techniques, laparoscopic right colectomy develops relatively slowly. This study was designed to compare the outcomes of laparoscopic right hemicolectomy (LRH) with open right hemicolectomy (ORH) in the treatment of colon carcinoma. METHODS: Between September 2000 and February 2003, 30 patients with colon cancer who underwent LRH were compared with 34 controls treated by ORH in the same period. All patients were evaluated with respect to surgery related complications, postoperative recovery, recurrence and metastasis rate, cost-effectiveness and survival. RESULTS: Among 30 LRH, 2 (6.7%) were converted to open procedure. No significant differences were observed in terms of mean operation time, blood loss, post-operative complications, and hospital cost between LRH and ORH groups. Mean time for bowel movement, hospital stay, and time to resum?early activity in the LRH group were significantly shorter than those in the ORH group (2.24±0.56 vs 3.25±1.29 d, 13.94?.5 vs 18.25±5.96 d, 3.94±1.64 vs 5.45±1.82 d respectively, P<0.05). As to the lymph node yield, the specimen length and total cost for operation and drugs, there was no significant difference between the two groups. Local recurrence rate and metachronous metastasis rate had no marked difference between the two groups. Cumulative survival probability at 40 mo in LRH group (76.50%) was not obviously different compared to the ORH group (74.04%). CONCLUSION: LRH in patients with colon cancer has statistically and clinically significant advantages over ORH. Thus, LRH can be regarded as a safe and effective procedure.展开更多
AIM:To explore the expression of Sp1 in gastric carcinoma as well as its association with other clinicopathologic features, and to evaluate the role of Spl as a prognostic indicator of gastric carcinoma. METHODS: By u...AIM:To explore the expression of Sp1 in gastric carcinoma as well as its association with other clinicopathologic features, and to evaluate the role of Spl as a prognostic indicator of gastric carcinoma. METHODS: By using immunohistochemistry, we examined the Spl expression patterns in 65 cases of human gastric cancer, and 40 normal gastric mucosa specimens. Simultaneously, the correlation between Spl expression and clinical outcome or clinicopathologic features was investigated. RESULTS: The percentage of Spl expression was 12.5% (5/40) in normal gastric mucosa, and the Spl protein was mainly expressed in the nuclei of cells located in the mucous neck region. In sharp contrast, strong Spl expression was detected in tumor cells, whereas no or faint Spl staining was detected in stromal cells and normal glandular cells surrounding the tumors. The expression rate of Spl in gastric cancer lesions was 53.85% (35/65). The medium survival duration in patients who had a tumor with negative, weak and strong Spl expressions was 1 700, 1 560 and 1 026 d, respectively (P<0.05). Spl protein expression was closely related to the depth of tumor infiltration (X2 = 13.223, P<0.01) and TNM stage (X2= 11.009, P<0.05), but had no relationship with the number of lymph nodes and Lauren's classification (P>0.05). Cox regression model for multivariate analysis revealed that high Spl expression (P<0.05) and advanced stage (P<0.01) were independent predictors of poor survival. CONCLUSION: Normal and malignant gastric tissues have unique Spl expression patterns. Spl might serve as an independent prognostic factor, by influencing the tumor infiltration and progression.展开更多
AIM: To evaluate the anti-tumor effects and possible involvement of anti-tumor immunity of electrochemotherapy (ECT) employing electroporation and bleomycin in human colon cancer xenografts in nude mice, and to establ...AIM: To evaluate the anti-tumor effects and possible involvement of anti-tumor immunity of electrochemotherapy (ECT) employing electroporation and bleomycin in human colon cancer xenografts in nude mice, and to establish the experimental basis for clinical application of ECT. METHODS: Forty nude mice, inoculated subcutaneously human colon cancer cell line LoVo for 3 wk, were allocated randomly into four groups: B+E+ (ECT), B+E- (administration of bleomycin alone), B-E+ (administration of electric pulses alone), and B-E- (no treatment). Tumor volumes were measured daily. The animals were killed on the 7* d, the weights of xenografts were measured, and histologies of tumors were evaluated. Cytotoxicity of spleen natural killer (NK) and lymphokine-activated killer (LAK) cells was then assessed by lactic dehydrogenase release assay. RESULTS: The mean tumor volume of group B+E+ was statistically different from the other three groups after the treatment (F= 36.80, P<0.01). There was one case of complete response, seven cases of partial response (PR) in group B+E+, one case of PR in group B+E- and group B-E+ respectively, and no response was observed in group B-E-. The difference of response between group B+E+ and the other three groups was statistically significant (X2 = 25.67,P<0.01). Histologically, extensive necrosis of tumor cells with considerable vascular damage and inflammatory cells infiltration were observed in group B+E+. There was no statistical difference between the cytotoxicity of NK and LAK cells in the four treatment groups. CONCLUSION: ECT significantly enhances the chemosensitivity and effects of chemotherapy in human colon cancer xenografts in nude mice, and could be a kind of novel treatment modality for human colon cancer. The generation of T-cell-dependent, tumor-specific immunity might be involved in the process of ECT.展开更多
AIM: To examine expression profile of gallbladder using microarray and to investigate the role of gallbladder in lipid homeostasis.METHODS: 33P-labelled cDNA derived from total RNA of gallbladder tissue was hybridized...AIM: To examine expression profile of gallbladder using microarray and to investigate the role of gallbladder in lipid homeostasis.METHODS: 33P-labelled cDNA derived from total RNA of gallbladder tissue was hybridized to a cDNA array representing 17 000 cDNA clusters. Genes with intensities ≥2 and variation ＜0.33 between two samples were considered as positive signals with subtraction of background chosen from an area where no cDNA was spotted. The average gray level of two gallbladders was adopted to analyze its bioinformatics. Identified target genes were confirmed by touch-down polymerase chain reaction and sequencing.RESULTS: A total of 11 047 genes expressed in normal gallbladder, which was more than that predicted by another author, and the first 10 genes highly expressed (high gray level in hybridization image), e.g. ARPC5 (2 225.88±90.46), LOC55972 (2 220.32±446.51) and SLC20A2 (1 865.21±98.02), were related to the function of smooth muscle contraction and material transport. Meanwhile, 149 lipid-related genes were expressed in the gallbladder, 89 of which were first identified (with gray level in hybridization image), e.g. FASN (11.42±2.62), APOD (92.61±8.90) and CYP21A2 (246.11±42.36), and they were involved in each step of lipid metabolism pathway. In addition, 19 of those 149 genes were gallstone candidate susceptibility genes (with gray level in hybridization image), e.g. HMGCR (10.98±0.31), NPC1 (34.88±12.12) and NR1H4 (16.8±0.65), which were previously thought to be expressed in the liver and/or intestine tissue only. CONCLUSION: Gallbladder expresses 11 047 genes and takes part in lipid homeostasis.展开更多
AIM:To provide hepatic protection through administration of doxorubicin before stop-flow chemotherapy (SFC) and to investigate the expression of heat shock protein 72 (HSP72) and role of nuclear factor kappa B (NF-κB...AIM:To provide hepatic protection through administration of doxorubicin before stop-flow chemotherapy (SFC) and to investigate the expression of heat shock protein 72 (HSP72) and role of nuclear factor kappa B (NF-κB) in this effect. METHODS: The hepatic preconditioning of doxorubicin was established in a porcine model by injection of doxorubicin (1 mg/kg) before SFC. The experimental animals were randomized into two groups: groups receiving doxorubicin (DOX) and normal saline (NS). Serial serum and tissue samples were taken from both groups to evaluate the protection of doxorubicin. Western blot and immunoprecipitation were applied to detect the expression of HSP72, NF-κB p65 protein, inhibitor κB-α (IκB-α) and phosphorylated IκB-α as well. The expression of tumor necrosis factor α (TNF-α) was estimated by semiquantitative RT-PCR. And the extent of the hepatic injury was estimated with the level of serum aminotransferases. RESULTS: An abundance production of HSP72 in porcine liver was observed after 24 h of intravenous administration of doxorubicin, but without any change in the expression of NF-κB p65 subunit in cytoplasm. NF-κB p65 subunit accumulated in nuclei at the end of SFC and reached its highest level at 30 min after the restoration of the abdominal circulation and decreased gradually during the 6 h after SFC in NS group, while there was little change in DOX group. There was also a slight decrease of IκB-α at 30 min after the restoration of the abdominal circulation in NS group accompanying with the appearance of phosphorylated IκB-α. The expression of TNF-α was significantly higher in NS group than that in DOX group (average 1.40±0.17 vs 0.62±0.22, P<0.01) at serial time points after SFC. Serum ALT and AST levels of NS group were higher after 24 h than those of DOX group (93.2±7.8 IU/L vs 53.3±13.9 IU/L, 217.0±29.4 IU/L vs 155.0±15.6 IU/L for ALT and AST respectively, P<0.05) and after 48 h than those of DOX group (66.6±18.1 IU/L vs 43.3±16.7 IU/L, 174.4±21.3 IU/L vs 125.7±10.5 IU/L for ALT and AST respectively, P<0.05). CONCLUSION: Doxorubicin renders the liver to be tolerant to the hepatic influence in SFC in a porcine model through the NF-κB/IκB-αpathway with the expression of HSP72.展开更多
Objective To study the inhibition effect on tumor angiogenesis and metastasis of angionstatin, which generated from human plasminogen. Methods Plasminogen was isolated metastasis of angio-from human plasma by Seph...Objective To study the inhibition effect on tumor angiogenesis and metastasis of angionstatin, which generated from human plasminogen. Methods Plasminogen was isolated metastasis of angio-from human plasma by Sepharose chromatography arm then catalyzed by elastase. Angiostatin was isolated by Sepharose 4B-Lysine chromatography. Nude mice model of metastatic gastric cancer was set up by intact tumor tissue implantation orthotopically. From the day of operation, mice received daily intraperitoneal injections of human angiostatin,intact plasminogen, or saline, respectively. 24μg ( 1.2mg/kg) of angiostatin or plasminogen was given on the day of operation, followed by a daily dose of 12μg( 0. 6mg /kg ) via intraperitoneal injection for three weeks.Ten weeks after implantation, mice were sacrificed and autopsied. Microvascular density was measured by im.munohistochemistry. Results Molecular weight of plasminogen isolated from plasma was 94KD. Plaanino-gen was catalyzed into two fragment peptides by elastase , which were 41～43KD and 51～53KD in molecular weight. Growth of the orthotopically implanted tumor was significantly reduced in size in the mice treated with angiostastin with an inhibition rate of 54.0%. Tumor metastasis to the liver and peritoneum was also signifi-cantly inhibited by angiostatin with inhibition rate of 61.9% and 55.6% respectively. The microvascular den-sity was also decreased significantly in the angiostatin treated mice. Conclusion Angiostatin may be generat-ed from plasma, and has inhibitory effect both on tumor growth and metastasis in nude mice model of human gastric cancer.展开更多
Background Since being reclassified by WHO in 1996, solid pseudopapillary tumour (SPT) of pancreas has been recognized as the internationally accepted name. Clinicians are lacking in knowledge of this rare disease so ...Background Since being reclassified by WHO in 1996, solid pseudopapillary tumour (SPT) of pancreas has been recognized as the internationally accepted name. Clinicians are lacking in knowledge of this rare disease so the misdiagnosis and inappropriate therapy are hard to avoid. The clinic data on 22 patients were summarized to study the misdiagnosis and treatment of a sample of SPTs. Methods Twenty-two female patients with SPT were studied retrospectively and divided into two groups, the misdiagnosed group and the correctly diagnosed one. The analyses were performed with Fisher test with accurate probability for categorical data, and Krusdal-Wallis test for ranked data.Results The rate of misdiagnosis in this sample was 45.5%. The misdiagnosed SPTs were apt to be the incomplete capsule ones (P=0.020), which resulted in obvious difficulties during operation (P=0.024). In the misdiagnosed SPT group, the medical expenses increased significantly (P=0.042), and the number of days in hospital greater than in correctly diagnosed group (P=0.041).Conclusions Although SPT has low malignancy with excellent prognosis after surgical treatment in most patients, the misdiagnosis of SPT increases the social and economic burdens on patients. It is important to analyse the causes of misdiagnosis.展开更多
文摘AIM:To study the expression of vascular endothelial growth factor C (VEGF-C) and chemokine receptor CCR7 in gastric carcinoma and to investigate their associations with lymph node metastasis of gastric carcinoma and their values in predicting lymph node metastasis.METHODS:The expression of VEGF-C and CCR7 in gastric carcinoma tissues obtained from 118 patients who underwent curative gastrectomy was examined by immunohistochemisty.Among these patients,39 patients underwent multi-slice spiral CT (MSCT) examination.RESULTS:VEGF-C and CCR7 were positively expressed in 52.5 and 53.4% of patients. VEGF-C expression was more frequently found in tumors with lymph node metastasis than those without it (P<0.001).VEGF-C expression was also closely related to lymphatic invasion (P<0.001), vascular invasion (P<0.01),and TNM stage (P<0.001). However,there was no significant correlation between VEGF-C expression and age at surgery, gender, tumor size, tumor location,Lauren classification,and depth of invasion. CCR7 expression was significantly higher in patients with lymph node metastasis compared with those without lymph node metastasis (P<0.001) and was also associated with tumorsize (P<0.01), depth of invasion (P<0.001), lymphatic invasion (P<0.001),and TNM stage (P<0.001).However,the presence of CCR7 had no correlation to age at surgery,gender, tumor location, Lauren classification,and vascular invasion. Among the 39 patients who underwent MSCT examination,only CCR7 expression was related to lymph node metastasis determined by MSCT (P<0.05).In the current retrospective study,the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of VEGF-C and CCR7 expression in the diagnosis of lymph node metastasis for patients with gastric carcinoma were 73.8%, 70.2%,72.6%,71.4% and 72.0%,and 82.0%,77.2%,79.4%,80.0% and 79.7%,respectively.After subdivision according to the combination of VEGF-C and CCR7 expression, receiver operating characteristic (ROC) analysis showed that the accuracy of the combined examination of VEGF-C and CCR7 expression in predicting lymph node metastasis was relatively high (area under ROC curve [Az]=0.83).CONCLUSION:The expression of VEGF-C and CCR7 is related to lymph node metastasis of gastric carcinoma and both of them may become new targets for the treatment of gastric carcinoma.Furthermore,the combined examination of VEGF-C and CCR7 expression in endoscopic biopsy specimens may be useful in predicting lymph node metastasis of gastric carcinoma and deciding the extent of surgical lymph node resection.
基金Supported by the Science Development Foundation of Shanghai,No. 95411902
文摘AIM: To study the interactive relationship of gallbladder motor function, plasma cholecystokinin (CCK) and cholecystokinin A receptor (CCK-R) of gallbladder in patients with cholesterol stone disease.METHODS: Gallbladder motility was studied by ultrasonography in 33 patients with gallbladder stone and 10 health subjects as controls. Plasma CCK concentration was measured by radioimmunoassay in fasting status (CCK-f) and in 30 min after lipid test meal (CCK-30).Radioligand method was employed to analyze the amount and activity of CCK-R from 33 gallstone patients having cholecystectomy and 8 persons without gallstone died of severe trauma as controls.RESULTS: The percentage of cholesterol in the gallstone composition was more than 70%. The cholesterol stone type was indicated for the patients with gallbladder stone in this study. Based on the criterion of gallbladder residual fraction of the control group, 33 gallstone patients were divided into two subgroups, contractor group (14 cases)and non-contractor group (19 cases), The concentration of CCK-30 was significantly higher in non-contractor group than that in both contractor group and control group (55.86±3.86 pmol/l vs 37.85±0.88 pmol/l and 37.95±0.74 pmol/L, P＜0.01), but there was no difference between contractor group and control group. Meanwhile no significant difference of the concentration of CCK-f could be observed among three groups. The amount of CCK-R was lower in non-contractor group than those in both control group and contractor group (10.27±0.94 fmol/mg vs24.59±2.39 fmol/mg and 22.66±0.55 fmol/mg, P＜0.01).The activity of CCK-R shown as KD in non-contractor group decreased compared to that in control group and contractor group. Only was the activity of CCK-R lower in contractor group than that in control group. The ejection fraction correlated closely with the amount of CCK-R (r = 0.9683,P＜0.01), and the concentration of CCK-30 correlated negatively with the amount of CCK-R closely (r = -0.9627,P＜0.01).CONCLUSION: The distinctive interactive relationship of gallbladder emptying, plasma CCK and CCK-R in gallbladder from this study suggested that the defect of CCK-R may be a key point leading to the impairment of gallbladder motor function and the pathogenesis of cholesterol gallstoneformation may differ in two subgroups of gallstone patient,gallbladder non-contractor group or contractor group.
文摘AIM: To investigate the clinical characteristics and prognostic factors of a consecutive series of patients with severe acute pancreatitis (SAP). METHODS: Clinical data of SAP patients admitted to our hospital from January 2003 to January 2004 were retrospectively reviewed. Collected data included the age, gender, etiology, length of hospitalization, APACHE Ⅱ score at admission, local and organ/systemic complications of the patients. RESULTS: Of the 268 acute pancreatitis patients, 94 developed SAP. The mean age of SAP patients was 52 years, the commonest etiology was cholelithiasis (45.7%), the mean length of hospitalization was 70 d, the mean score of APACHE Ⅱ was 7.7. Fifty-four percent of the patients developed necrosis, 25% abscess, 58% organ/systemic failure. A total of 23.4% (22/94) of the SAP patients died. Respiratory failure was the most common organ dysfunction (90.9%) in deceased SAP patients, followed by cardiovascular failure (86.4%), renal failure (50.0%). In the SAP patients, 90.9% (20/22) developed multiple organ/systemic failures. There were significant differences in age, length of hospitalization, APACHE Ⅱ score and incidences of respiratory failure, renal failure, cardiovascular failure and hematological failure between deceased SAP patients and survived SAP patients. By multivariate logistic regression analysis, independent prognostic factors for mortality were respiratory failure, cardiovascular failure and renal failure. CONCLUSION: SAP patients are characterized by advanced age, high APACHE Ⅱ score, organ failure and their death is mainly due to multiple organ/systemic failures. In patients with SAP, respiratory, cardiovascular and renal failures can predict the fatal outcome and more attention should be paid to their clinical evaluation.
文摘AIM:To determine the expression levels of three metabolic enzymes of fluoropyrimidines:thymidylate synthase (TS),thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) in seven human gastrointestinal cancer cell lines,and to compare the enzyme levels with the sensitivity to 5-fluorouracil (5-FU) and 5-fluoro-2′-deoxyuridine (FdUrd).METHODS:TS,TP and DPD mRNA levels were assessed by semi-quantitative RT-PCR,TP and DPD protein contents were measured by ELISA. Fifty percent inhibitory concentrations of growth (IC50),representing the sensitivity to drugs,were determined by MTT assay.RESULTS:IC50 values ranged from 1.28 to 12.26μM for 5-FU,and from 5.02 to 24.21μM for FdUrd,respectively.Cell lines with lower DPD mRNA and protein levels tended to be more sensitive to 5-FU (P<0.05), but neither TS nor TP correlated with 5-FU IC50 (P>0.05).Only TS mRNA level was sharply related with FdUrd sensitivity (P<0.05),but TP and DPD were not (P>0.05).A correlation was found between mRNA and protein levels of DPD (P<0.05),but not TP (P<0.05).CONCLUSION:DPD and TS enzyme levels may be useful indicators in predicting the antitumor activity of 5-FU or FdUrd,respectively.
基金Supported by a grant from Shanghai Science and Technology Committee in China,No.954119027
文摘AIM: To investigate the relationship between gallbladder stone disease (GSD) and single nucleotide polymorphisms of cholesterol 7α-hydroxylase (CYP7A) gene promoter,apolipoprotein (APO) B gene exon 26, APOEgene exon 4 or microsatellite polymorphism of low density lipoprotein receptor (LDLR) gene exon 18.METHODS: Genotypes of CYP7A, APOB, APOE and LDLR genes were determined in 105 patients with GSD diagnosed by B-mode ultrasonography and 274 control subjects.Serum lipids were analyzed with HITACHI 7060 automaic biochemical analyzer.RESULTS: Body mass index (BMI) was significantly higher in patients with GSD (24.47±3.09) than in controls (23.50±2.16).Plasma total cholesterol was lower in patients with GSD (4.66±0.92 mmol/L) than in controls (4.91±0.96 mmol/L),P<0.01 after adjusted for age, sex and BMI. The significantly higher frequency of A allele of CYP7A gene polymorphism and X+ allele of APOBgene polymorphism was seen in GSD patients. Percentages of A allele in patients and controls were 62.86% and 54.38% (P <0.05) and those of X+ allele 8.57% and 4.01% (P<0.01). Subjects with A allele had significantly lower plasma total cholesterol and LDL cholesterol than subjects with CC homozygote. In a multiple variable logistic regression model, the BMI (OR=1.13, 95% CI: 1.05-1.22), A allele (OR=1.48, 95% CI: 1.05-2.09) and X+ allele (OR=2.28, 95% CI: 1.14-4.59) were positively associated with GSD (P <0.05). Plasma total cholesterol (OR=0.69, 95% CI: 0.64-0.74) was negatively related to GSD (P<0.05).CONCLUSION: With an association analysis, it was determined that A allele of CYP7A gene and X+ allele of APOB gene might be considered as risk genes for GSD. These alleles are related with differences of serum lipids among subjects.Multiple-variable logistic regression model analysis showed that besides BMI, GSD was affected by polygenetic factors.But the mechanism for these two alleles responsible for GSD requires further investigations.
基金Supported by National Natural Science Foundation of China,No.30170915,Health Ministry of China,No.9802292,and Shanghai Medical Development Foundation from the Health Bureau of Shanghai,No.983008
文摘AIM:To investigate the immunotherapeutic potential of vaccine consisting of dendritic cells (DCs) pulsed with total RNA from MFC gastric cancer cells.METHODS:DCs were prepared from the spleens of strain 615 mice by magnetic cell sorting (MACS). Alter culture for 24h, DCs were pulsed with total RNA from MFC gastric cancer cells. Mice of one group were immunized with tumor RNA pulsed DC (RNA/DC) at the dosage of 1×10^6 on d 14 and 7 by s c inoculation before tumor implantation. Mice of another group were immunized with unpulsed DC (UDC) at the same dosage on days as the RNA/DC group. The third group of control mice was untreated. On d 0, all the mice were challenged with s c injections of 5×10^5 MFC gastric cancer cells.Alter inoculation, the mice were monitored closely with respect to tumor growth. Activities of NK cells in PBL and splenocytes and CTL were tested.RESULTS:On d 21 alter tumor cell inoculation, the mice of control group manifested the largest tumors with volume at a mean of 2.6323±1.1435cm^3, followed by the UDC and RNA/DC groups with mean volumes at 0.7536±0.3659cm^3 and 0.3688±0.6571cm^3, respectively. The activities of NK cells in PBL and splenocytes in RNA/DC group were 66.2% and 65.4%, respectively, higher than that in the control group. The tumor specific CTL activity in RNA/DC group was 49.5%, higher than that in the control group.CONCLUSION:The tumor vaccine with DCs pulsed with total RNA from gastric cancer cells possesses the ability to stimulate tumor specific CTL activity and to establish antitumor immunity when administered in vivo.
基金Health Ministry Scientific and Research Foundation of China,No.98-1-312
文摘AIM: Cancer gene therapy has received more and moreattentions in the recent decade. Various systems of genetherapy for cancer have been developed. One of the mostpromising choices is the suicide gene. The product ofthymidine kinase (TK) gene can convert ganciclovir (GCV)to phosphorylated GCV, which inhibits the synthesis of cellDNA, and then induces the cells to death. Cytolines play animportant role in anti-tumor immunity. This experiment wasdesigned to combine theTK gene and mIL-2/mGM-CSFgenes to treat gastric cancer, and was expected to producea marked anti-tumor effect.METHODS: TK gene was constructed into the retroviralvector pLxSN, and the mIL-2 and mGM-CSF genes wereinserted into the eukaryotic expressing vector pIRES. Thegastric cancer cells were transfected by retroviral serum thatwas harvested from the package cells. In vitro study, thetransfected gastric cancer cells were maintained in the GCV-contained medium, to assay the cell killing effect andbystander effect. In vivo experiment, retroviral serum andcytokines plasmid were transfected into tumor-bearing mice,to observe the changes of tumor volumes and survival ofthe mice.RESULTS: In vitro experiment, 20 % TK gene transducedcells could cause 70-80 % of total cells to death. In vivoresults showed that there was no treatment effect in controlgroup and TK/GCV could inhibit the tumor growth. Thestrongest anti-tumor effect was shown in TK+mIL-2+mGM-CSF group. The pathologic examination showed necrosis ofthe cancer in the treated groups.CONCLUSION: TK/GCV can kill tumor cells and inhibit thetumor growth in vivo IL-2 and GM-CSF strongly enhancethe anti-tumor effect. Through the retrovirus and liposomemethods, the suicide gene and cytokine genes are allexpressed in the tissues.
文摘AIM: To evaluate the effect of reactive oxygen species suchas hydrogen peroxide on the progression of human coloncancer.METHODS: Human colon carcinoma cell lines, LS174T andHCT8, were treated respectively with 10- 5,10- 7 or 10- 9 mol@L- 1 hydrogen peroxide for 24h, and co-cultured with humanendothelial cell line ECV-304. The migration of ECV-304induced by cancer cells was calculated and the expressionlevel of vascular endothelial growth factor in cancer cellswas determined by RT-PCR analysis and ELISA.Dactinomycin of 1.5mg@ L-1 which could block transcriptionof cancer cells was applied to observe the effects of H2O2 ontranscriptional activity and the relative half-life of VEGFmRNA. Finally, to evaluate the effect H2O2 on NF-κB activityin colon cancer cells, NF-κB in cytoplasm and nucleus of thecells were detected with FITC-tagged antibody and itspresence in the nucleus (Fn ) Vs cytoplasm ( Fc ) wasmonitored by measuring the green fluorescence integratedover the nucleus by laser scanning cytometry(LSC).RESULTS: Exogenouse hydrogen peroxide of lowconcentration increased the migration of endothelial cellsinduced by colon cancer cells. When cancer cells weretreated with 10-5 mol@ L-1 H2O2, the migration number ofendothelial cells induced by LS174T cells was 203 ± 70, andthe number induced by HCT8 cells was 145 ± 65. The twovalues were significantly higher than those treated with otherconcentrations of h2O2 ( P ＜ 0.01 ). The expression ofvascular endothelial growth factor in cancer cells, whichcould be blocked by dactinomycin, were increased to acertain degree, while the relative half-life of VEGF mRNAwas not prolonged after treatment with hydrogen peroxide.The activity of NF-κB in colon cells rose after the cells wereexposed to hydrogen peroxide for 24h. The Fn values inHCT8 cells were 91 ± 13 (0 mol@ L- 1 h2O2) and 149 ± 40( 10-5mol@L-1 h2O2) ( P＜ 0.05), in LS174T cells were 127 ± 35(0mol@L-1 H2O2) and 192± 11(10-5 mol@L-1 h2O2) (P＜ 0.05).lt is similar to the case of VEGF expression in cancer cells.CONCLUSION: Hydrogen peroxide increases vascularendothelial growth factor expression in colon cancer cells,and it is likely that reactive oxygen species such ashydrogen peroxide facilitates the development of coloncancer.
基金Supported by Science and Technology Development Foundation of Shanghai,No. 024119106
文摘AIM: Laparoscopic surgery, especially laparoscopic rectal surgery, for colorectal cancer has been developed considerably. However, due to relatively complicated anatomy and high requirements for surgery techniques, laparoscopic right colectomy develops relatively slowly. This study was designed to compare the outcomes of laparoscopic right hemicolectomy (LRH) with open right hemicolectomy (ORH) in the treatment of colon carcinoma. METHODS: Between September 2000 and February 2003, 30 patients with colon cancer who underwent LRH were compared with 34 controls treated by ORH in the same period. All patients were evaluated with respect to surgery related complications, postoperative recovery, recurrence and metastasis rate, cost-effectiveness and survival. RESULTS: Among 30 LRH, 2 (6.7%) were converted to open procedure. No significant differences were observed in terms of mean operation time, blood loss, post-operative complications, and hospital cost between LRH and ORH groups. Mean time for bowel movement, hospital stay, and time to resum?early activity in the LRH group were significantly shorter than those in the ORH group (2.24±0.56 vs 3.25±1.29 d, 13.94?.5 vs 18.25±5.96 d, 3.94±1.64 vs 5.45±1.82 d respectively, P<0.05). As to the lymph node yield, the specimen length and total cost for operation and drugs, there was no significant difference between the two groups. Local recurrence rate and metachronous metastasis rate had no marked difference between the two groups. Cumulative survival probability at 40 mo in LRH group (76.50%) was not obviously different compared to the ORH group (74.04%). CONCLUSION: LRH in patients with colon cancer has statistically and clinically significant advantages over ORH. Thus, LRH can be regarded as a safe and effective procedure.
基金Supported by the "211" Project sponsored by Ministry of Education,China
文摘AIM:To explore the expression of Sp1 in gastric carcinoma as well as its association with other clinicopathologic features, and to evaluate the role of Spl as a prognostic indicator of gastric carcinoma. METHODS: By using immunohistochemistry, we examined the Spl expression patterns in 65 cases of human gastric cancer, and 40 normal gastric mucosa specimens. Simultaneously, the correlation between Spl expression and clinical outcome or clinicopathologic features was investigated. RESULTS: The percentage of Spl expression was 12.5% (5/40) in normal gastric mucosa, and the Spl protein was mainly expressed in the nuclei of cells located in the mucous neck region. In sharp contrast, strong Spl expression was detected in tumor cells, whereas no or faint Spl staining was detected in stromal cells and normal glandular cells surrounding the tumors. The expression rate of Spl in gastric cancer lesions was 53.85% (35/65). The medium survival duration in patients who had a tumor with negative, weak and strong Spl expressions was 1 700, 1 560 and 1 026 d, respectively (P<0.05). Spl protein expression was closely related to the depth of tumor infiltration (X2 = 13.223, P<0.01) and TNM stage (X2= 11.009, P<0.05), but had no relationship with the number of lymph nodes and Lauren's classification (P>0.05). Cox regression model for multivariate analysis revealed that high Spl expression (P<0.05) and advanced stage (P<0.01) were independent predictors of poor survival. CONCLUSION: Normal and malignant gastric tissues have unique Spl expression patterns. Spl might serve as an independent prognostic factor, by influencing the tumor infiltration and progression.
基金Supported by the Grant from Science and Technology Development of Shanghai, No. 00440
文摘AIM: To evaluate the anti-tumor effects and possible involvement of anti-tumor immunity of electrochemotherapy (ECT) employing electroporation and bleomycin in human colon cancer xenografts in nude mice, and to establish the experimental basis for clinical application of ECT. METHODS: Forty nude mice, inoculated subcutaneously human colon cancer cell line LoVo for 3 wk, were allocated randomly into four groups: B+E+ (ECT), B+E- (administration of bleomycin alone), B-E+ (administration of electric pulses alone), and B-E- (no treatment). Tumor volumes were measured daily. The animals were killed on the 7* d, the weights of xenografts were measured, and histologies of tumors were evaluated. Cytotoxicity of spleen natural killer (NK) and lymphokine-activated killer (LAK) cells was then assessed by lactic dehydrogenase release assay. RESULTS: The mean tumor volume of group B+E+ was statistically different from the other three groups after the treatment (F= 36.80, P<0.01). There was one case of complete response, seven cases of partial response (PR) in group B+E+, one case of PR in group B+E- and group B-E+ respectively, and no response was observed in group B-E-. The difference of response between group B+E+ and the other three groups was statistically significant (X2 = 25.67,P<0.01). Histologically, extensive necrosis of tumor cells with considerable vascular damage and inflammatory cells infiltration were observed in group B+E+. There was no statistical difference between the cytotoxicity of NK and LAK cells in the four treatment groups. CONCLUSION: ECT significantly enhances the chemosensitivity and effects of chemotherapy in human colon cancer xenografts in nude mice, and could be a kind of novel treatment modality for human colon cancer. The generation of T-cell-dependent, tumor-specific immunity might be involved in the process of ECT.
基金Supported by the National Natural Science Foundation of China,No. 30271272, and the Foundation of Chinese National Human Genome Center at Shanghai, No. CHCS-99M-06
文摘AIM: To examine expression profile of gallbladder using microarray and to investigate the role of gallbladder in lipid homeostasis.METHODS: 33P-labelled cDNA derived from total RNA of gallbladder tissue was hybridized to a cDNA array representing 17 000 cDNA clusters. Genes with intensities ≥2 and variation ＜0.33 between two samples were considered as positive signals with subtraction of background chosen from an area where no cDNA was spotted. The average gray level of two gallbladders was adopted to analyze its bioinformatics. Identified target genes were confirmed by touch-down polymerase chain reaction and sequencing.RESULTS: A total of 11 047 genes expressed in normal gallbladder, which was more than that predicted by another author, and the first 10 genes highly expressed (high gray level in hybridization image), e.g. ARPC5 (2 225.88±90.46), LOC55972 (2 220.32±446.51) and SLC20A2 (1 865.21±98.02), were related to the function of smooth muscle contraction and material transport. Meanwhile, 149 lipid-related genes were expressed in the gallbladder, 89 of which were first identified (with gray level in hybridization image), e.g. FASN (11.42±2.62), APOD (92.61±8.90) and CYP21A2 (246.11±42.36), and they were involved in each step of lipid metabolism pathway. In addition, 19 of those 149 genes were gallstone candidate susceptibility genes (with gray level in hybridization image), e.g. HMGCR (10.98±0.31), NPC1 (34.88±12.12) and NR1H4 (16.8±0.65), which were previously thought to be expressed in the liver and/or intestine tissue only. CONCLUSION: Gallbladder expresses 11 047 genes and takes part in lipid homeostasis.
基金Supported by the Shanghai Bureau of Health, 99ZDII003
文摘AIM:To provide hepatic protection through administration of doxorubicin before stop-flow chemotherapy (SFC) and to investigate the expression of heat shock protein 72 (HSP72) and role of nuclear factor kappa B (NF-κB) in this effect. METHODS: The hepatic preconditioning of doxorubicin was established in a porcine model by injection of doxorubicin (1 mg/kg) before SFC. The experimental animals were randomized into two groups: groups receiving doxorubicin (DOX) and normal saline (NS). Serial serum and tissue samples were taken from both groups to evaluate the protection of doxorubicin. Western blot and immunoprecipitation were applied to detect the expression of HSP72, NF-κB p65 protein, inhibitor κB-α (IκB-α) and phosphorylated IκB-α as well. The expression of tumor necrosis factor α (TNF-α) was estimated by semiquantitative RT-PCR. And the extent of the hepatic injury was estimated with the level of serum aminotransferases. RESULTS: An abundance production of HSP72 in porcine liver was observed after 24 h of intravenous administration of doxorubicin, but without any change in the expression of NF-κB p65 subunit in cytoplasm. NF-κB p65 subunit accumulated in nuclei at the end of SFC and reached its highest level at 30 min after the restoration of the abdominal circulation and decreased gradually during the 6 h after SFC in NS group, while there was little change in DOX group. There was also a slight decrease of IκB-α at 30 min after the restoration of the abdominal circulation in NS group accompanying with the appearance of phosphorylated IκB-α. The expression of TNF-α was significantly higher in NS group than that in DOX group (average 1.40±0.17 vs 0.62±0.22, P<0.01) at serial time points after SFC. Serum ALT and AST levels of NS group were higher after 24 h than those of DOX group (93.2±7.8 IU/L vs 53.3±13.9 IU/L, 217.0±29.4 IU/L vs 155.0±15.6 IU/L for ALT and AST respectively, P<0.05) and after 48 h than those of DOX group (66.6±18.1 IU/L vs 43.3±16.7 IU/L, 174.4±21.3 IU/L vs 125.7±10.5 IU/L for ALT and AST respectively, P<0.05). CONCLUSION: Doxorubicin renders the liver to be tolerant to the hepatic influence in SFC in a porcine model through the NF-κB/IκB-αpathway with the expression of HSP72.
文摘Objective To study the inhibition effect on tumor angiogenesis and metastasis of angionstatin, which generated from human plasminogen. Methods Plasminogen was isolated metastasis of angio-from human plasma by Sepharose chromatography arm then catalyzed by elastase. Angiostatin was isolated by Sepharose 4B-Lysine chromatography. Nude mice model of metastatic gastric cancer was set up by intact tumor tissue implantation orthotopically. From the day of operation, mice received daily intraperitoneal injections of human angiostatin,intact plasminogen, or saline, respectively. 24μg ( 1.2mg/kg) of angiostatin or plasminogen was given on the day of operation, followed by a daily dose of 12μg( 0. 6mg /kg ) via intraperitoneal injection for three weeks.Ten weeks after implantation, mice were sacrificed and autopsied. Microvascular density was measured by im.munohistochemistry. Results Molecular weight of plasminogen isolated from plasma was 94KD. Plaanino-gen was catalyzed into two fragment peptides by elastase , which were 41～43KD and 51～53KD in molecular weight. Growth of the orthotopically implanted tumor was significantly reduced in size in the mice treated with angiostastin with an inhibition rate of 54.0%. Tumor metastasis to the liver and peritoneum was also signifi-cantly inhibited by angiostatin with inhibition rate of 61.9% and 55.6% respectively. The microvascular den-sity was also decreased significantly in the angiostatin treated mice. Conclusion Angiostatin may be generat-ed from plasma, and has inhibitory effect both on tumor growth and metastasis in nude mice model of human gastric cancer.
文摘Background Since being reclassified by WHO in 1996, solid pseudopapillary tumour (SPT) of pancreas has been recognized as the internationally accepted name. Clinicians are lacking in knowledge of this rare disease so the misdiagnosis and inappropriate therapy are hard to avoid. The clinic data on 22 patients were summarized to study the misdiagnosis and treatment of a sample of SPTs. Methods Twenty-two female patients with SPT were studied retrospectively and divided into two groups, the misdiagnosed group and the correctly diagnosed one. The analyses were performed with Fisher test with accurate probability for categorical data, and Krusdal-Wallis test for ranked data.Results The rate of misdiagnosis in this sample was 45.5%. The misdiagnosed SPTs were apt to be the incomplete capsule ones (P=0.020), which resulted in obvious difficulties during operation (P=0.024). In the misdiagnosed SPT group, the medical expenses increased significantly (P=0.042), and the number of days in hospital greater than in correctly diagnosed group (P=0.041).Conclusions Although SPT has low malignancy with excellent prognosis after surgical treatment in most patients, the misdiagnosis of SPT increases the social and economic burdens on patients. It is important to analyse the causes of misdiagnosis.
