Search

Objective To determine the clinicopathological significance of RhoC expression in human ovarian cancer and its effect on the expression of vascular endothelial growth factor (VEGF), Rho-associated coiled-coil-forming kinase (ROCK), and metal matrix proteinases (MMPs). Methods Tissue samples from normal ovaries, benign ovarian tumors, and epithelial ovarian cancer were collected. The mRNA and protein expression levels of RhoC, ROCK-I, VEGF, and MMP9 were assessed using reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blot, and compared to the clinicopathological characteristics of the sample of origin using the Pearson method of correlation analysis. Small interfering RNA (siRNA) was also used to target RhoC expression in the OVCAR3 and CaOV3 ovarian cancer cell lines, after which cell invasion and migration assays were performed, and the expression of ROCK-I, VEGF, and MMP9 was evaluated. Results The expression levels of RhoC, ROCK-I, VEGF, and MMP9 mRNA and protein were significantly higher in ovarian cancer, showing a correlation with clinical stage but not histological type. RhoC expression was positively correlated with ROCK-I, VEGF, and MMP9 expression. Decreased RhoC expression in siRNA-targeted cells inhibited their ability to invade and migrate, as well as inhibiting ROCK-I, VEGF, and MMP9 expression. Conclusion The expression level of RhoC is correlated to clinical stage and vascularization in ovarian cancer.

To clarify the clinicopathological features and prognosis, and to evaluate the rational surgical therapy for superficial spreading early gastric cancer (EGC).Superficial spreading EGC was defined as tumor invading the mucosa or submucosa layer with the diameteror =50 mm, and common EGC as tumor with diameter50 mm. The clinicopathological data of 62 patients with superficial spreading EGC and 224 patients with common EGC were collected and analyzed retrospectively.No significant differences in age, sex, differentiation, depth of invasion, growth manner, vessel involvement and lymph node metastasis were found between superficial spreading and common EGC(P0.05), but significant differences were seen in tumor site, macroscopic type, scope of gastrectomy and lymphadenectomy between the two groups(P0.05). Superficial spreading EGC located in the middle or the whole of stomach accounted for 45.2%, and presented mixed macroscopic type more frequently(48.4%), such as II(C+II(A 5 cases, II(C+II(B 9 cases and II(C+III( 16 cases, and more total gastrectomy and more D(2) lymphadenectomy(16.1% and 40.3% respectively) were executed, compared with common EGC. Due to cancer residual,two patients underwent enlargement of the scope of gastrectomy,two underwent total gastrectomy and one underwent operation again after surgery. The 5-, 10-year tumor-free survival rates for superficial spreading EGC were 98.4% and 91.4%, while 97.0% and 91.2% for the common EGC (chi(2)=1.16,P=0.282).Special clinicopathological features can be seen in superficial spreading EGC, and the lesion is superficial and extensive. Prevention of cancer residual in remnant stomach is the key point and D(2) lymphadenectomy is suitable enough for operation.

To summarize the features of metastasis in different lymph node groups (from 1 to 16 groups) in gastric cancer patients treated by total gastrectomy, and evaluate their clinical significance in lymph node dissection.The data of 73 gastric cancer patients with total gastrectomy and lymph node dissection from January 2004 to April 2006 were analyzed retrospectively. The lymph nodes were divided into 16 groups according to the 13(th) edition of gastric cancer treatment guideline of JGCA (The Japan Gastric Cancer Association). The metastatic rate and degree of dissected lymph nodes in these patients were compared.The metastatic rates of lymph node groups in these patients from lower to higher were as follows: group 15, 13/16, 14v, 12, 10, 9, 11, 8, 2, 6/7, 5, 1, 4, 3. The lowest was the 15(th) group lymph nodes (1.4%), the highest was the 3rd group (65.8%), with a statistically significant difference between those two groups (P0.01). The metastatic degrees of the lymph node groups from lower to higher were as follows: 13, 16, 1, 7, 6, 5, 12, 4, 11, 8, 2, 15, 9, 3, 10, 14v. There was a statistically significant difference between the lowest group of lymph node (13(th) group, 10.7%) and the highest (14v(th), 56.3%, P0.01).In the radical total gastrectomy for patients with gastric cancer, it is suggested that the regional lymph nodes with higher metastatic rate should be resected necessarily, and the group with a higher metastatic degree should be dissected completely. If the result of sentinel lymph node biopsy in the 3(rd) or 14v(th) group is negative, the operation extent can be reduced. If positive, it should be extended. When the biopsy result in the 13(th) or 16(th) is positive, palliative operation may be indicated. However, if the biopsy result is negative in the 13(th) or 16(th), but positive in the 14v(th) group, extended operation is indicated.

To analyze the clinicopathologic features and prognosis of young patients with gastric cancer by comparing with older patients.The clinicopathologic data of 157 younger adults (age,/= 40 years) with gastric cancer and 1761 cases of elder gastric cancer patients (age,40 years) was analyzed and compared retrospectively. All of the 1918 patients were surgically treated between January 1980 and December 2000.The rates of poorly differentiation, diffusive growth, Borrmann 4 type, whole-stomach invasion were significantly higher in younger cases than those in the elder counterparts (P0.05), especially in young female patients. The rate of early gastric cancer was significantly higher in young patients than that in older patients (P0.05), especially in young male patients. There was significant difference between the survival rate of younger male cases (median survival, 35 months) and younger female cases (median survival, 19 months) (P = 0.0219), but no significant difference was found between elder male and elder female (median survival, 26 vs. 30 months). TNM stage, operative curability, gross type were independent predictive factors of survival for younger patients.Younger female gastric cancer patients tends to have worse prognosis than older patients, while younger male patients have better prognosis due to higher percentage of early gastric cancer when diagnosed. Pathologic staging and operative curability are the independent predictive factors of survival for younger patients.

To investigate the effect of lymphadenectomy adjacent to inferior mesenteric artery root on the prognosis of rectal cancer.Clinicopathological data of 260 cases with rectal cancer undergone radical operation were analyzed retrospectively. The patients were divided into two groups. Group D(2): the lymph nodes adjacent to mesenteric artery root were not excised (n=188). Group D(3): the lymph nodes adjacent to mesenteric artery root were excised (n=72). Prognosis of two groups was compared during the follow-up period.In group D(2), the 1-, 3-, 5-year total survival rates (TS) were 97.3%, 87.2% and 77.1%, and tumor-free survival rates (TFS) were 93.1%, 83.0% and 76.8% respectively. In group D(3 ), the 1-, 3-, 5-year total survival rates (TS) were 94.4%, 79.2% and 73.6%, and tumor-free survival rates (TFS) were 86.1%, 76.4% and 71.0% respectively. The differences of TS and TFS between two groups were not significant according to Kaplan-Meier analysis (P0.05). Multivariate analysis revealed that the excision of lymph nodes adjacent to mesenteric artery root was not statistically correlated with the recurrence, metastasis and survival time after radical operation of rectal cancer.Excision of lymph nodes adjacent to inferior mesenteric artery root has no significant impact on prognosis and it is unnecessary in the radical operation of rectal cancer.

To investigate the inhibitory effect of siRNA on heparanase expression and invasion ability gastric cancer cells.A heparanase mRNA-targeting double-stranded siRNA was designed with the bioinformatics technology. Human gastric cancer cells of the line SGC7901 were cultured and transfected with the siRNA of the concentrations of 5, 10, 20, and 40 nmol/L respectively. Forty-eight hours later RT-PCR and Western blotting were applied to detect the mRNA and protein expression of heparanase. Millicell chamber assay was performed to detect the invasion ability of the SGC7901 cells. Blank control group and negative control group were set.The mRNA expression level of the cells transfected with the siRNA of the concentrations 20 nmol/L and 40 nmol/L were 0.207 +/- 0.095 and 0.200 +/- 0.085 respectively, both significantly lower than that of the control group (0.60 +/- 0.09, both P0.05). Western blotting showed that the protein expression of heparanase of the different siRNA subgroups were all decreased dose-dependently; and no heparanase band was seen in the 40 nmol/L subgroup. The invasion rate of the siRNA group was significantly lower than that of the control group with a mean inhibition rate of (61 +/- 36)%.RNAi inhibits the expression of heparanase and the invasion ability of human gastric cancer cells. Heparanase may be a new target in treatment of gastric cancer's metastasis.

To evaluate the efficacy of different modes of lymphadenectomies in the surgical treatment for early gastric cancer (EGC) in order to provide evidence for reasonable selection of lymphadenectomy for the ECG.The clinicopathological data of 325 EGC patients were reviewed. The lymph node metastasis (LNM) rate of different levels and the efficiency of different modes of lymphadenectomies were analyzed, and the correlation between LNM and clinicopathological features were analyzed either.The overall LNM rate in the EGC was 14.8%, and the overall LNM degree was 3.0%. In the ECG located in the lower third of the stomach, the rate of first level nodal involvement was 14.5%, which was detected in the No. 3, No. 4, No. 5, No. 6 station of draining lymph nodes for the stomach; and it was 6.9% in the second level lymph nodes which were seen in the No.7 and No. 8a station. There was rare metastasis in the No. 1, No. 9, No. 11p, No. 12a and No. 14v station. In the ECG located in the middle third of the stomach, the rate of first level nodal involvement was 13.8% , which was found in the No. 1, No. 3, No. 5 and No. 6 station, and it was 6.9% in the second level nodes only seen in the No. 7 and No. 8a station. The LNM rate was significantly higher in the lesion larger than 3.0 cm in diameter, or having submucosa invasion, or being poorly differentiated, or with lymphatic involvement than that of the lesion smaller than 3.0 cm in diameter, without mucosa invasion or lymphatic involvement, or being well differentiated (P0.05).Based on our data, it is suggested that the D1 or D1 + lymphadenectomy of the No. 7 station of lymph node may be suitable for the lesion smaller than 1.0 cm in diameter or only located in the mucosa; the D1 + lymphadenectomy of the No. 7 and No. 8a station of lymph nodes may be suitable for the lesion larger than 1.0 cm in diameter, or being depressed type or with submucosa invasion in the middle and/or lower third of the stomach. For the lesion larger than 3.0 cm in diameter, the No. 1 and No. 9 station lymph node should be dissected. It is also suggested that regular D2/ D3 lymphadenectomy should be avoided in the surgical treatment for the early gastric cancer whenever the tumor situation is suitable.

Andrographolide, an active component isolated from the Chinese official herbal Andrographis paniculata, has recently been reported to have anticancer activity. However the molecular mechanism responsible for its anticancer action has not been fully defined. In this study, we investigated the effect of andrographolide on the adhesion of gastric cancer cells to the activated endothelial cells and the expression of some cell adhesion molecules.Human endothelial cells were preincubated with andrographolide for 6 h and then incubated with the cytokine tumor necrosis factor for 4 h. Endothelial surface expression of E-selectin was evaluated by flow cytometry, immunostaining and ELISA. Further, we investigated E-selectin mRNA expression by RT-PCR. Surface expression of sialyl Lewis(X) of three gastric cancer cell lines (SGC7901, MGC803, BGC823) and a normal gastric epithelial cell line GES-1 was evaluated by flow cytometry and immunostaining. Adherence of CFSE-labeled gastric cancer cells and GES-1 cells to endothelial cell monolayers was then determined.Andrographolide significantly reduced E-selectin expression of activated endothelial cells, and inhibited the E-selectin expression on mRNA level. Three gastric cancer cell lines expressed high levels of sialyl Lewis(X), whereas GES-1 did not. Andrographolide also significantly decreased gastric cancer cells adherence to stimulated endothelial cells. The inhibition of E-selectin expression corresponded to the reduction of tumor cell adherence. The effects of andrographolide on tumor adhesion were almost nullified by pre-incubation with E-selectin and sialyl Lewis(X) antibody.These findings demonstrate that andrographolide suppresses the adhesion of gastric cancer cells which express high level sialyl Lewis(X) to human vascular endothelial cells by blocking E-selectin expression and, thus, may represent a candidate therapeutic agent for cancer.

To objectively evaluate the practical significance of different extended surgeries in early gastric cancer(EGC) patients, and to choose reasonable gastrectomies and lymphadenectomies.A total of 217 EGC patients were investigated undergone normalized D2 or above extended surgery and their clinicopathological data were recorded in detail. The efficiency of the extended lymphadenectomies, complications and operation causes were analyzed, and the correlation between the group 2 lymph node metastasis (LNM) and clinicopathological factors were assessed, too.There was no nodal involvement in the No.5 and No.6 lymph nodes among the total gastrectomy in the upper third of the stomach, neither was in the No.10, 11p and 11d lymph nodes among the combined splenectomy, and neither was in the No.15 lymph nodes among the combined transverse mesocolon resection in the lower third of the stomach. There was no distant nodal involvement in the EGC. Above all, most of them were mistaken for advanced gastric cancer preoperatively and intraoperatively, the operation time was longer and the blood loss was more during operation. Among the resected nodes of group 2 in the lower third of the stomach, metastasis was not found in the No.11p, 12a and 14v lymph nodes. The rate of the No.7 and 8a nodal involvement in the submucosa cancer was higher than that in the mucosa cancer(P0.05) and so did the No.7 in the lymphatic penetration positive(P0.001). The No.1 and No.13 nodal involvement were only seen in the high risk cases, such as submucosa cancer, the lesion diameter more than 3.0 cm, depressed type and lymphatic involvement.It is not necessary to execute total gastrectomy in the upper third of the stomach, combined organ resection (such as splenectomy, transverse mesocolon resection), and distant lymph node dissection in the EGC. In the lower third of the stomach, the No.11p 12a and 14v lymph nodes shouldn't be dissected. With respect to the high risk nodal involvement cases in the lower third of the stomach, the No.1 lymph nodes should be dissected and so does the No.13 lymph nodes if it's tumefied. It is the key point of reasonable operation to exactly diagnose the EGC before and during the surgery.

To evaluate the status of lymph node metastasis (LNM) and reasonable procedure in early gastric cancer (EGC).Two hundred and ninety-two patients with histologically proven mucosal or submucosal gastric cancer who underwent gastrectomy/lymphadenectomy were included in this study. The numbers of total dissected lymph node were all above 15 in all patients. The clinical characteristics, pathologic features, and LNM were assessed by univariate and multivariate analysis.LNM were observed in 43 of 292 cases (14.7%), and 6.4% in mucosal lesions and 22.4% in submucosal lesions. The LNM was identified in 12.7% at the first level, 7.2% at the second level and 0.34% at the third level. The LNM frequency was found in the 6, 3, 4, 1, 5 lymph node (from high to low) consequently at the first level and the 7, 8a lymph node at the second level. The EGC with nodal involvement and the tumors2.0 cm in diameter were all depressed type. The diameter of elevated type with LNM was no less than 3.0 cm in this series. The depth of invasion and lymphatic vessel involvement were independent influencing factors in LNM on multivariate analysis (P0.05).Less extensive surgery might be considered for the elevated type EGC, and tumorsor= 2.0 cm in diameter and mucosal lesions. Gastrectomy with D1 plus No.7, 8a lymphadenectomy might be carried out in the depressed type/or= 3.0 cm in diameter, or in the elevated type/3.0 cm in diameter. With respect to the depressed type EGC/tumors3.0 cm in diameter, gastrectomy with D2 lymphadenectomy is proper.

To explore the feasibility of transfecting DHFR (human double-mutant dihydrofolate reductase) gene into mouse bone marrow cells and the effect of resistance to high dose MTX chemotherapy.After DHFR gene was transfected into mouse bone marrow cells with retroviral vector, the cells were treated with methotrexate (MTX) and then CFU-GM (granulocyte-macrophage colony-forming unit) assay was performed. Peripheral blood leucocytes and platelets, body weight and survival rate were observed. After treatment with high dose MTX, the expression of drug resistance gene was checked by RT-PCR in the transfected bone marrow cells.SFG-F/S-NeoR gene-transfected mice bone marrow cells yielded drug-resistance colonies to MTX (donor mice: 15.8%, recipient mice: 18.0%, control: 0) The peripheral blood leucocytes and platelets, body weight recovered gradually and the survival rate was 83.3% at the 40th day, while 0 in controls in gene transfected mice after large dose MTX treatment. RT-PCR of transgenic mouse marrow cells showed the band of F/S gene (400 bp).DHFR gene can not only be integrated and expressed in bone marrow cells but also improve their drug-resistence to MTX.

To investigate the effects of cimetidine on the proliferation, apoptosis, adhesion, and invasion of human gastric cancer cells.Human gastric cancer cells of the line SGC-790 were cultured. Cimetidine of the concentrations of different concentrations was added into the culture fluid. The survival rate of the cells was calculated. Flow cytometry was used to examine the distribution of cell cycle and apoptosis of the cells. Fluorescence staining was used to observe the morphology of the apoptotic cells. The adhesion of the cells was measured by MTT method and their invasion ability was tested with Millicell chambers.The proliferation of the AGC-7901 cells was inhibited by cimetidine of the concentration of 0.5 to 5 mmol/L time and dose-dependently. Cimetidine of the concentrations of 0.5 - 10 mmol/L retarded the SGC-7901 cells at the stage G(0)/G(1) and increased the number of apoptotic cells dose-dependently (P0.05). Cimetidine of the non-toxic concentrations 250, 125, and 62.5 micrommol/L decreased the adhesion and invasion of the SGC-7901 cells to the extracellular matrix.Capable of inducing apoptosis and cell cycle arrest, cimetidine is helpful in treatment of cancers of gastric cancer.

Chromosome 8p21-p23 harbors tumor suppressor gene(s) implicated in multiple types of cancers. To investigate the involvement of the gene(s) in the carcinogenesis of adenocarcinoma of gastric cardia, loss of heterozygosity (LOH) for microsatellite markers at chromosome 8p21-p23 was examined. Laser capture microdissection (LCM) was used to obtain homogeneous tumor cells from 19 surgical specimens. Subsequently, genomic DNA extracted from the LCM-captured cells was amplified by multiple displacement amplification. Each tumor was assessed for allelic loss using 13 microsatellite markers. An overall LOH frequency of 63.2% (12/19) was observed and the LOH frequency for individual markers varied from 25% to 55.6%. One common deleted region of about 1.2 Mb (8p22GGAA-8p22ATCT) was defined. Our data indicated that the tumor suppressor gene at chromosome 8p22 might play an important role in the development of adenocarcinoma of gastric cardia.

To study the expression and significance of interleukin-8 (IL-8), cyclooxygenase-2 (COX-2) and trefoil family factor 1 (TFF1) in the remnant stomach mucosa.Patients after gastrectomy were examined by upper gastrointestinal endoscopy. Biopsy specimens were obtained from stoma and the greater curvature of the upper corpus to be assessed for Hp (by H.E. and Giemsa staining) and conduct real-time semi-quantitative PCR. mRNA was extracted from the biopsy specimens to determine the IL-8, COX-2 and TFF1 gene mRNA levels by real-time PCR method.In the stoma, COX-2 level in Hp-positive patients was significantly higher than that in Hp-negative patients, but the difference of IL-8 levels between them was not significant. In the corpus, IL-8 and COX-2 levels in Hp-positive patients were significantly higher than those in Hp-negative patients. In Hp-negative patients, IL-8 and COX-2 levels in the stoma were significantly higher in B II anastomosis than in B I anastomosis cases; COX-2 level in the stoma was significantly higher in B II anastomosis than in B I anastomosis cases, but the difference of IL-8 levels between them was not significant. TFF1 level in the remnant stomach mucosa showed no significant difference between Hp-positive and Hp-negative patients.Hp infection and bile reflux are important risk factors for the secondary stomach carcinogenesis. Expression of IL-8 and COX-2 in the remnant stomach mucosa is related to the risk of secondary stomach carcinogenesis. The relationship between the TFF1 expression and secondary stomach carcinogenesis in the remnant stomach mucosa is still unclear and should further be studied.

to investigate the relationship between metastatic rate and amount of lymph nodes and prognosis of gastric cancer.The clinical data of 672 patients of gastric cancer with 22.7 regional metastatic lymph nodes on average were analyzed retrospectively. The patients were divided into 3 groups according to the TNM staging system (1997, 5th edition): pN0 (without metastatic lymph node), pN1 group (with 1-6 metastatic lymph nodes), pN2 group (with 7-15 metastatic lymph nodes), and pN3 group (with more than 215 metastatic lymph nodes), or into 4 groups according to the lymph node metastasis rate: rN0 group (without metastasis of lymph node), rN1 group (with a lymph node metastasis rate 010%), rN2 group (with a lymph node metastasis rate of 10%-30%), and rN3 group with a lymph node metastasis rate30%). The 5-year survival rates of different groups in these 2 classification systems and the relationship between metastatic rate and amount of lymph nodes and prognosis of gastric cancer were analyzed.According to the lymph node metastasis grading system there were 93 patients in the pN0 group, 201 in the pN1 group, 108 in the pN2 group, and 32 in the pN3 group with the 5-year survival rates of 90.3%, 65.2%, 37.0%, and 18.9% respectively (chi(2) = 12.56, P0.01). According to the lymph node metastasis system 93 patients were in the rN0 group, 183 in the rN1 group, 92 in the rN2 group, and 66 in the rN3 group, with the 5-year survival rates of 90.3%, 69.9%, 45.7%, and 10.6% respectively (chi(2) = 14.38, P0.01). If the cases in the same pN group were re-divided according to the lymph node metastasis rate, there were significant differences in the 5-year survival rate among the new rN groups (chi(2) = 5.26, P = 0.21), and there was a great difference in the distribution of prognosis among different groups. However, if the cases in the same group rN group were re-divided according to the amounts of metastatic lymph nodes, there was no significant difference in 5-year survival rate among different new pN groups (chi(2) = 0.14, P = 0.932). Multivariate COX model analysis showed that the influencing factors of the prognosis were rN grading, pN grading, infiltration depth of cancer, gross type, and size of the cancer, arranged according to the degree of closeness (P0.05); lymph nose metastasis rate was closely related with the size, infiltration depth, and gross type of the cancer (P0.05). In the same metastasis rate, the 5-year survivals rate decreased along with the increase of the amount of metastatic lymph nodes, and vice versa (chi(2) = 8.22, P = 0.004; chi(2) = 4.52, P = 0.033).The staging system based on metastatic lymph node rate is more predictable the number of metastatic lymph nodes for the prognosis of gastric cancer.

To study the clinical value of computer aided detection (CAD) in direct full-field digital mammography.480 digital mammograms of 120 patients with pathologically proved breast cancer were randomly selected and interpreted by an expert radiologist without knowing the pathologic diagnosis, then analyzed by CAD system, and finally read by the radiologist again to make the final diagnosis. The sensitivity for breast cancer detection of the radiologist in diagnosis of breast cancer was calculated before and after using CAD. Data of the breast density, diameters of mass and calcification, being spiculate or not, and pathology of each case were reviewed to determine how they impacted on the improvement of sensitivity with the assistance of CAD.The sensitivity for breast cancer detection by the radiologist before using CAD was 74.2%, significantly lower than that after using CAD (89.2%, chi(2) = 37.628, P = 0.000). Breast density (chi(2) = 7.352, P = 0.007), diameter of calcification (chi(2) = 10.549, P = 0.014) and pathologic pattern (chi(2) = 13.751, P = 0.003) were highly correlated with the improvement of sensitivity. Use of CAD might overcome type 1 missed diagnosis, and not type 2 missed diagnosis.Computer-aided detection helps radiologist improve their diagnostic accuracy of breast cancer, however, is not capable of differentiating benign from malignant lesions.

To explore the feasibility of transferring fusion gene of dihydrofolate reductase (DHFR) gene and cytidine deaminase (CD) gene into mouse bone marrow cells in order to observe the drug resistance of high dose methotrexate (MTX) and cytosine arabinoside (Ara-C) in the bone marrow cells and to improve the tolerance of myelosuppression following combination chemotherapy.Human double-mutant dihydrofolate reductase-cytidine deaminase fusion gene was transferred into two mice bone marrow cells by retroviral vector. Resistant colony-forming unit granulocyte-macrophage (CFU-GM) assays were performed in mouse bone marrow cells by retroviral infection and after treatment by drugs (Ara-C, MTX, and Ara-C + MTX). DNA was extracted from mouse bone marrow cells. The expression of drug resistant genes in mouse bone marrow cells after transferring by retroviral vector was checked by polymerase chain reaction (PCR).Bone marrow cells after coculture with the retroviral producer cells transduced with the genes (SFG-F/S-CD) showed the drug resistance colonies yield (Colony formation after exposure to Ara-C, MTX and Ara-C + MTX were 56%, 22% and 14%, respectively) and the increase in drug resistant to both MTX and Ara-C (P0.005). Expression of DHFR and CD gene in extracted DNA of transfected mice were demonstrated by PCR.Double drug resistant gene can not only integrate and co-express in mice bone marrow cells but also increase the drug resistance to MTX and Ara-C.

To evaluate bromodeoxyuridine/DNA (BrdUrd/DNA) double-parametric flow cytometric analysis in detection of gastric carcinoma and to study the correlations of cellular BrdUrd labeling index (LI), G(2)/M phase fraction (G(2)/MPF) and DNA content, with lymphatic and vascular invasion and prognosis.Sixty cases of fresh tumor samples were examined by BrdUrd/DNA double-parametric flow cytometry.BrdUrd LI and G(2)/MPF values were both significantly higher in patients with lymphatic invasion than those without invasion (P0.01). There was statistically significant difference between the 5-year survival rates in cases with and without lymphatic invasion (P0.01). Both BrdUrd LI and G(2)/MPF values were significantly higher in patients with lymph node metastasis than those in cases without metastasis (P0.01). There was a significant difference in 5-year survival rates between patients with and without lymph node metastases. The incidence of lymph node metastasis was significantly higher in aneuploid carcinoma (P0.05), and the patients with aneuploidy had significantly poor prognosis. BrdUrd LI was significantly higher in patients with more than 5 metastatic lymph nodes than those with 1-4 metastatic lymph nodes (P0.05) and no metastasis (P0.01). G(2)/MPF values in cases with more than 5 and 1-4 metastatic lymph nodes were higher than that in cases with no metastasis (P0.01 and P0.05, respectively). There were significant differences in 5-year survival rates among patients with no lymph node metastasis, 1-4 nodes metastasis and more than 5 nodes metastasis. G(2)/MPF values were significantly higher in patients with and without vascular invasion (P0.01).Correlations exist among cellular BrdUrd LI, G(2)/MPF, DNA content and lymphatic involvement, vascular invasions and prognosis in gastric carcinoma.

Microcalcifications play a very important role in detection of breast cancer, especially early stage breast cancer. However, ambiguity still exists in understanding the relationship between radiological and pathological characteristics of microcalcifications. The definitive indication of a biopsy has not been established. The purpose of this study is to evaluate the relationship of classification of breast microcalcifications using full-field digital mammography to the pathological characteristics.For all the women an open biopsy had been conducted. One hundred and three mammographs showing clustered microcalcifications from 98 consecutive patients were reviewed along with their pathological records. To investigate the value of each criterion for the detection of cancer, univariate and multivariate analyses were performed on the entire sample and then on morphological subgroups.Pathological examination showed 67 malignant lesions (65.05%) and 36 benign lesions (34.95%). In the univariate analysis, four radiological variables were significant: morphological type (P = 0.001), complicated by a mass (P = 0.002), number of microcalcifications per cluster (P = 0.02) and linear or triangular distribution of clusters (P = 0.009). In the multivariate analysis, two criteria remained significant: morphological type (P0.001) and complicated by a mass (P = 0.001). The percentage of malignancy was 37.0%, 60.0%, 78.8%, and 88.9%, respectively, for type 2 (regularly punctiform), type 3 (dusty), type 4 (irregularly punctiform) and type 5 (vermicular) microcalcifications (Le Gal's classification). The malignancy was 78.6% for microcalcifications complicated by a mass and 48.9% without a mass. The difference was significant (P0.05). The relationship between morphological types of microcalcifications and the pathological characteristics was also studied. In subgroups, type 3 (dusty) microcalcifications complicated by a mass (P = 0.001) or with the number of microcalcifications more than 10 (P = 0.024); and type 2 (regularly punctiform) with a diameter of the area over 20 mm (P = 0.024) or complicated by a mass (P = 0.025) were statistically significant as criteria for malignant tumour.Most cases of microcalcifications of type 4 or 5; type 3 complicated by a mass or with the number of microcalcifications more than 10; type 2 complicated by a mass or with a diameter of the area over 20 mm; are indicative of cancer. Open biopsy is recommended to acquire definitive pathological diagnosis for these cases. For the remainder of the morphological types, stereotaxic biopsy or followup should be considered.

STK15 (Serine/Threonine protein kinase 15) is a serine/threonine kinase encoding gene, whose overexpression in mammalian cells leads to centrosome amplification, chromosomal instability, and oncogenic transformation. 91A--T, a single nucleotide polymorphism (SNP) in exon 3 of STK15, has recently been shown to be associated with human cancer susceptibility. Within exon 3 of STK15, there are three nonsynonymous SNPs: 91 A--T, 169G--A and 311C--T. We have determined STK15 genotypes and haplotypes composed of 91A--T and 169G--A by PCR-RFLP in a randomly sampled cohort of 193 normal individuals from Northeast China. DNA samples from all individuals were subjected to first round of PCR using a pair of specific primers. For the subsequent nested PCR,a mismatch forward primer, which could introduce an EcoR I restriction site to the 91A allele, was included. The nested PCR products were digested with the restriction endonucleases EcoRI and AccII. The double restriction digests were separated by polyacrylamide gel electrophoresis. Three haplotypes,91A-169G, 91T-169A and 91T-169G, were detected and their frequencies were 68.65%, 10.88% and 20.47%, respectively. Six genotypes composed of the above three haplotypes were found,and their frequencies were 91A-169G/91 A-169G (46.11%), 91A-169G/91 T-169A (14.51%), 91A-169G/91T-169G (30.57%), 91T-169G/91T-169G (3.11%), 91T-169G/91T-169A (4.15%), 91T-169A/91T-169A (1.55%). Whereas no 91A-169A haplotype was detected in all individuals examined in the current study, linkage disequilibrium (LD) between the two SNPs was found.

To explore the relationship between helicobacter pylori (Hp) infection and expression of interleukin-8 (IL-8) in the remnant stomach mucosa.Fifty-eight patients with gastrectomy were examined by upper gastrointestinal endoscopy. Biopsy specimens were obtained from stoma and the greater curvature of the upper corpus to detect Hp infection (by H.E. and Giemsa staining) and IL-8 expression (by real-time semi-quantitative RT-PCR).Hp infection was detected in 65.5% (38/58)of the remnant stomach mucosa. In 34 patients with Billroth I (BI) anastomosis, IL-8 in corpus was significantly higher in Hp-positive patients than that in Hp-negative patients (0.11+/-0.07 vs 0.02+/-0.01, P0.05). In 24 patients with Billroth II (BII) anastomosis, IL-8 in stoma and corpus in Hp-positive patients was significantly higher than that in Hp-negative patients (P0.05).Hp infection induces IL-8 expression in remnant stomach mucosa. In corpus, IL-8 mRNA expression is mainly related with Hp infection, while in stoma, IL-8 mRNA expressions may be related with bile reflux besides Hp infection.

To study the expressions and the significance of interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) in the remnant stomach.Fifty-eight patients with gastrectomy were examined by upper gastrointestinal endoscopy. Two biopsy specimens were obtained from the stoma and the upper corpus gastric mucosa in the remnant stomach. mRNA was extracted from biopsy specimens to measure the IL-8 and COX-2 gene mRNA levels by real-time PCR method.IL-8 and COX-2 levels were higher in stoma than in corpus, IL-8 levels in BI anastomosis were significantly higher in stoma than in corpus (P0.05). In Hp-negative patients, IL-8 and COX-2 levels in stoma were significantly higher in BII anastomosis than in BI anastomosis (P0.05). In Hp-positive patients, IL-8 and COX-2 levels in stoma showed no significant differences between BII anastomosis and BI anastomosis. In corpus, IL-8 and COX-2 levels in Hp-positive patients were significantly higher than those in Hp-negative patients, (P0.05), including in BI anastomosis and in BII anastomosis.The risk of the secondary stomach carcinogenesis in stoma after distal gastrectomy is higher than that in corpus; The types of anastomosis may influence the risk for the secondary stomach carcinogenesis in the remnant stomach mucosa.

To screen and analyze the important associated genes in different stages of gastric cancer.Using suppression subtractive hybridization (SSH) to screen differentially expressed genes; detecting the expression of genes in different stages of gastric cancer with dot blot hybridization; and verifying the results with semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR).Twenty-six differentially expressed gene fragments were obtained by means of SSH. Among them,24 were known genes, 1 was a new expressed sequence tags(EST), and 1 was a hypothetical gene. The results of dot blot hybridization demonstrated that the expressions of Annexin A2, RPS29, RPS12 etc. in dysplasia were higher than those in normal mucosa; the expressions of RPS12 etc.in early cancer were higher than those in normal mucosa;the expressions of cytochromosome C oxidase II, ferritin light chain, RPS12 etc. in advanced gastric cancer and lymph node metastases were consistently higher than those in normal mucosa. The expression of proteasome 26S subunit gene in advanced gastric cancer was higher than that in normal mucosa. The expression of RPS12 was consistently higher in different stages of gastric cancer. It was demonstrated by RT-PCR that the expression of RPS12 in gastric cancer was higher than that in normal mucosa.The authors have identified some important genes that might be involved in the carcinogenesis and progression of gastric cancer, and RPS12 may play more important roles in gastric cancer.

In order to explore the correlation of the abnormalities of tumor suppressor gene APC with the carcinogenesis and progression of gastric cancer. The I1307K mutation of APC gene in gastric cancer was analysed using Amplification Refractory Mutation System PCR(ARMS ,PCR),also the expression of APC protein in gastric cancer of different stages was detected by immunohistochemical method. We found that there wasn't I1307K mutation of APC gene in 62 cases of blood samples of susceptible population in high incidence areas of gastric cancer and 45 cases of gastric cancer tissues. The positive rates of APC protein in gastric cancer (both early and progressive gastric cancer) were significantly lower than that in normal mucosa,the positive rates of APC protein in progressive gastric cancer were significantly lower than that in early gastric cancer,the positive rates of APC protein in gastric cancer with lymph node metastasis were significantly lower than that in gastric cancer without lymph node metastasis. So it was thought that there might be no correlation between the I1307K mutation of APC gene and carcinogenesis of gastric cancer in China,but the decreased expression of APC protein was closely related to the carcinogenesis,progression and lymph node metastasis of gastric cancer.

To optimize and screen the most suitable target gene length,concentration and printing solution in cDNA microarray, housekeeping genes, such as beta actin and GAPDH, were selected as targets and hepatitis B virus gene as negative control. The RT-PCR primers that spanned at least one intron and whose products were at between 189 bp and 1078 bp were designed with primer premier 5.0, so did the hepatitis B virus gene PCR primer. After polymerase chain reaction, the products were purified with ethanol and dissolved in 3xSSC, 50% DMSO and 0.5 mol/L carbonate buffer(pH=9.0)respectively. The concentrations of target genes were adjusted at 0.5 microg/microL, 1.0 microg/microL and 1.5 microg/microL. The hybridization signals had a good specificity. No signal showed in either negative control (HBV) or blank control (printing solution only). There was no significant difference in target gene lengths. The P value of beta actin (189 bp,491 bp,974 bp) and GAPDH (227 bp,552 bp,1078 bp) was 0.378 and 0.866 respectively. There was no significant difference among concentrations(P=0.648),too. However, the higher the concentration was, the stronger the signals would be. Among the three kinds of printing solution, 50% DMSO was the best (P=0.0001), while the other two had no difference by multi-comparison (P=0.142). The target gene at length between 200 bp and 1000 bp has got the same hybridization signals.50% DMSO printing solution and the target gene concentration of 0.5 microg/microl are suitable for good hybridization.

This study was to evaluate bivariate bromodeoxyuridine (BrdUrd)/DNA flow cytometric analysis in detection of gastric carcinoma and to study the relations of cellular BrdUrd labeling indices (LI), G2/M-phase fraction (G2/MPF) and DNA ploidy pattern to lymphatic involvement, venous invasion and prognosis.Fresh tumor samples from 60 patients with gastric carcinoma were analyzed by bivariate BrdUrd/DNA flow cytometry. The results were correlated with lymphatic vessel invasion, lymphatic node metastasis, the number of metastatic lymphatic nodes, and venous invasion. Propidium iodide (PI) was used as a fluorescent probe for total cellular DNA, and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescent-labeled goat anti-mouse antibody was used as a second antibody. S-phase fractions were measured by in vitro BrdUrd labeling, and DNA ploidy and G2/MPF were also measured. Comparison of survival was performed with the log-rank test, the Chi-square test for qualitative data, and Student's t test for quantu data.BrdUrd LI and G2/MPF values were significantly higher in tumors with lymphatic vessel invasion than in those without invasion respectively (P0.01); the patients who had tumors with lymphatic vessel invasion showed a significantly poor prognosis (P0.01). Both BrdUrd LI and G2/MPF values were significantly higher in tumors with lymphatic node metastasis than in those without metastasis (P0.01). A statistical significant difference was noted in the 5-year survival rates between the patients with lymph node metastasis and those without metastasis. Compared with diploid carcinoma, the incidence of lymph node metastasis was significantly higher in aneuploid carcinoma (P0.05), and the patients with aneuploid carcinoma showed a significantly poor prognosis (P0.05). BrdUrd LI was significantly higher in patients with more than 5 metastatic lymph nodes than those with 1 - 4 metastatic lymph nodes (P0.05) and those without metastasis (P0.01). G2/MPF values in those patients either with more than 5 metastatic lymph nodes or 1 - 4 metastatic lymph nodes were higher than those without metastasis (P0.01 and P0.05). A statistical significance was seen in the 5-year survival rates among the patients with no metastatic lymph node, 1 - 4 metastatic nodes and more than 5 metastatic nodes (P0.01). G2/MPF values were significantly higher in patients with venous invasion than in those without invasion (P0.01).Positive correlations exist between cellular BrdUrd LI, G2/MPF with lymphatic involvement and prognosis, and DNA aneuploid with lymphatic involvement and prognosis. The same was true between G2/MPF value and venous invasion in gastric carcinoma.

To investigate the inhibitory effect of hyaluronic acid (HA), hyaluronidase (Hase) and arg-gly-asp tripeptide (RGD) on adhesion and invasion of human gastric cancer cell line SGC7901.Expression of CD44 and integrin beta1 protein on cell surface was determined by indirect fluorescence. After SGC7901 cells were treated with HA, Hase and RGD alone or in various combinations, their adhesion and invasion to ECM were measured by MTT and Boyden chamber method. Cell morphology was also observed.Expression of CD44 and integrin beta1 protein on cell surface was detected in human gastric cancer cell line SGC7901. Hase or RGD alone could block the adhesion and invasion of SGC7901 cells to ECM in contrast to the control (P0.001, P0.05); their blocking effect was stronger than HA (P0.05). The inhibitory effect of Hase + HA or a combination of the three agents was stronger than any single agent (P0.001). Morphologically, the untreated cells adhered onto the matrigel had spread out presenting a fibroblast feature with variously shaped pseudopods, while the treated ones were kept round with relatively fewer pseudopods.HA, Hase and RGD can inhibit the adhesion and invasion of SGC7901 cells expressing functional CD44 and integrin beta1 protein to ECM, and a combination of the three agents may achieve the best inhibitory effect.

To construct cDNA subtracted libraries from gastric dysplasia and further screen differentially expressed genes.Relatively pure dysplasia and normal tissue were procured by manual microdissection, and amplified by cDNA-PCR, which was used to carry on for suppression subtractive hybridization (SSH). Subtracted cDNA fragments were linked with vector, cloned, screened, sequenced, and made homologous search. Differentially expressed fragments were verified by dot hybridization.Two subtracted cDNA libraries were constructed. Among 26 sequenced clones, 15 fragments corresponded to known genes, 3 fragments were known EST and 8 fragments were unknown EST (GenBank BQ164614-BQ164616, BQ291516-BQ291520). Fifteen fragments were verified to be differentially expressed in gastric dysplasia.Subtracted cDNA libraries from gastric dysplasia are constructed using combination of microdissection-cDNA PCR and SSH setup in our laboratory. Some fragments have been screened and verified to help to search for novel associated genes with gastric carcinogenesis.

Aminopeptidase N has been identified as the cellular receptor for human coronavirus HCoV-229E and was a putative receptor for the spike glycoprotein encoded by the SARS-associated coronavirus (SARS-CoV). We report here identification of 9 single nucleotide polymorphisms (SNPs) in ANPEP, encoding human aminopeptidase N, in Chinese. All ANPEP exons and their flanking intronic sequences were amplified from unrelated normal individuals by polymerase chain reaction (PCR) and screened using denaturing high-performance liquid chromatography (DHPLC). Nine SNPs were revealed after direct sequencing of PCR amplified fragments which showed changes of DHPLC chromatogram. Four of these polymorphisms, T321M(962CT), S651L(1952CT), S752N(2255GA) and G764R(2290GA), were non-synonymous; the remaining exonic synonymous and intronic ones were T795T(2385CT), IVS7 + 17GA, IVS14-16AG, IVS17 + 12CG and IVS17 + 44CT. Our data may be useful for studies to investigate the role of host genetic factors in SARS pathogenesis, especially for identifying SARS-susceptible and/or anti-SARS alleles.

To study the relationship between hRad17 mRNA expression and clinicopathologic factors and lymph node metastasis of gastric cancer, and to assess the significance of predicting the extent of lymph node metastasis and prognosis.hRad17 mRNA expression was examined in matched primary lesions, normal gastric mucosa and lymph node metastatic lesions among 52 gastric cancer patients by reverse transcription polymerase chain reaction (RT-PCR), polyacrylamide gel electrophoresis (PAGE) and silver stain with the relation between hRad17 mRNA expression and clinicopathologic factors analyzed. At the same time, hRad17 mRNA expressions in 5 gastric benign lesions and SGC7901 gastric carcinoma cell lines were also examined.The primary tumor samples (88.4% positive) showed a significantly higher level of hRad17 expression compared with matched normal tissue (76.9% positive) (P = 0.014), so did the lymph node metastatic samples (94.2% positive) (P = 0.001). The hRad17 mRNA expression showed a low level in benign lesions, but very high in SGC7901 cell line. The hRad17 mRNA expression showed a higher level in patients with the number of lymph node metastasis above 15 than below 15 (P = 0.02), so did the diffused growth than the mass-like growth (P = 0.04).The method of PAGE and silver stain can improve the sensitivity of RT-PCR. The degree of lymph node metastasis and invasiveness of carcinoma cells are more serious in cases with hRad17 mRNA overexpression, and extensive lymph node dissection should be carried out for these patients. Examination of hRad17 expression by RT-PCR before surgery is indicated to arrive at an optimum treatment scheme and to estimate the prognosis.

Gastric cancer is the second most common cause of cancer-related death world-wide. The carcinogenesis, progression and metastasis of gastric cancer involve many genetic events, and go through multi...

AIM: To determine the effect of three digestive tract reconstruction procedures on pouch function, after radical surgery undertaken because of gastric cancer, as assessed by radionuclide dynamic imaging. METHODS: As a measure of the reservoir function, with a designed diet containing technetium-99m (99mTc), the emptying time of the gastric substitute was evaluated using a 99mTc-labeled solid test meal. Immediately after the meal, the patient was placed in front of a γ camera in a supine position and the radioactivity was measured over the whole abdomen every minute. A frame image was obtained. The emptying sequences were recorded by the microprocessor and then stored on a computer disk. According to a computer processing system, the half-emptying actual curve and the fitting curve of food containing isotope in the detected region were depicted, and the half-emptying actual curves of the three reconstruction procedures were directly compared. RESULTS: Of the three reconstruction procedures, the half-emptying time of food containing isotope in the Dual Braun type esophagojejunal anastomosis procedure (51.86 ± 6.43 min) was far closer to normal, significantly better than that of the proximal gastrectomy orthotopic reconstruction (30.07 ± 15.77 min, P = 0.002) and P type esophagojejunal anastomosis (27.88 ± 6.07 min, P = 0.001) methods. The half-emptying actual curve and fitting curves for the Dual Braun type esophagojejunal anastomosis were fairly similar while those of the proximal gastrectomy orthotopic reconstruction and P type esophagojejunal anastomosis were obviously separated, which indicated bad food conservation in the reconstructed pouches. CONCLUSION: Dual Braun type esophagojejunal anastomosis is the most useful of the three procedures for improving food accommodation in patients with a pouch and can retard evacuation of solid food from the reconstructed pouch.

AIM: To detect the presence of inducible nitric oxide synthase (iNOS), nitrotyrosine (NT) and apoptosis in gastric adenocarcinomas and their possible correlations with the clinicopathological characteristics and prognosis of gastric adenocarcinoma. METHODS: Sixty-six specimens of gastric adenocarcinoma and corresponding adjacent normal gastric tissues were studied. Immunohistochemistry was employed to localize iNOS and NT protein and an immunohistochemical scoring system was used. The occurrence of apoptotic cell death (apoptotic index [AI]) was analyzed by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labeling (TUNEL) method. RESULTS: Results showed that iNOS expression was detected at an intermediate or high level in 41 of 66 (62%) specimens of gastric adenocarcinoma. NT expression was 58%. Neither of them was found in the normal gastric tissues; there were significant positive correlations among iNOS expression, NT expression and AI. Many clinicopathologic characteristics of gastric adenocarcinoma, such as tumor size, depth of invasion, lymph node metastasis and TNM staging, were related to iNOS and NT expressions (P

AIM:To explore molecular mechanism of gastric carcinogenesis, we screened associated genes of gastric dysplasia and further investigated their expression in gastric carcinomas with different stages. METHODS:Relatively pure dysplasia and normal tissue were procured by manual microdissection, amplified by cDNAPCR, and then used to carry out forward (dysplasia as tester, normal tissue as driver) and reverse (normal tissue as tester, dysplasia as driver) SSH. Subtracted cDNA fragments were cloned into vector, screened, sequenced, and made homologous analysis. The expression of differentially expressed fragments was detected and verified by Dot hybridization and reverse transcription-PCR. RESULTS:Two subtracted cDNA libraries were constructed. Twenty-one of 26 sequenced clones were verified to be expressed differentially. It was noted that differentiall expressions of 4 genes (P125,cytochrome c oxidase subunit I, meprin A,acidic calponin)were detected simultaneously in dysplasia, early cancer and advanced cancer.