Prognostic value and clinical correlations of18-fluorodeoxyglucose metabolism quantifiers in gastriccancer

AIM： To investigate the correlations of pre-treatmentpositron emission tomography-computer tomography（PET-CT） metabolic quantifiers with clinical data ofunstratified gastric cancer （GC） patients.METHODS： Forty PET-CT scans utilising 18-fluorodeoxyglucosein patients who received no prior treatmentwere analysed. Analysis involved measurements ofmaximum and mean standardised uptake volumes（SUV）, coefficient of variation （COV）, metabolictumour volumes and total lesion glycolysis of differentthresholds above which the tumor volumes wereidentified. The threshold values were： SUV absolutevalue of 2.5, 30% of SUVmax, 40% of SUVmax,and liver uptake-based （marked 2.5, 30, 40 and liv,respectively）. Clinical variables such as age, sex,clinical stage, performance index, weight loss, tumorhistological type and grade, and CEA and CA19.9 levelswere included in survival analysis. Patients receivedvarious treatment modalities appropriate to theirdisease stage and the outcome was defined by time tometastasis （TTM） and overall survival （OS）. Clinical andmetabolic parameters were evaluated by analysis of variance, receiver operating characteristics, univariateKaplan-Meier, and multivariate Cox models. P 〈 0.05was considered statistically significant.RESULTS： Significant differences were observedbetween initially disseminated and non-disseminatedpatients in mean SUV （6.05 vs 4.13, P = 0.008）, TLG2.5（802 cm3 vs 226 cm3; P = 0.031）, and TLG30 （436 cm3vs 247 cm3, P = 0.018）. Higher COV was associatedwith poor tumour differentiation （0.47 for G3 vs0.28 for G1 and G2; P = 0.03）. MTV2.5 was positivelycorrelated to patient weight loss （〈 5%, 5%-10%and 〉 10%： 40.4 cm3 vs 123.6 cm3 vs 181.8 cm3,respectively, P = 0.003）. In multivariate Cox analysis,TLG30 was prognostic for OS （HR = 1.001, 95%CI：1.0009-1.0017; P = 0.047） for the whole group ofpatients. In the same model yet only including patientswithout initial disease dissemination TLG30 （HR = 1.009,95%CI： 1.003-1.014; P = 0.004） and MTV2.5 （HR = 1.02,95%CI： 1.002-1.036; P = 0.025） were prognostic forOS; for TTM TLG30 was the only significant prognosticvariable （HR = 1.006, 95%CI： 1.001-1.012; P = 0.02）.CONCLUSION： PET-CT in GC may represent a valuablediagnostic and prognostic tool that requires furtherevaluation in highly standardised environments such asrandomised clinical trials.

The characteristics of DNA repair synthesis induced by DNA polymerase β in hepatoma cells after γ rays irradiation

AIM To investigate the effects of DNA repair synthesis induced by DNA polymerase β in hepatoma cells after γ ray irradiation. METHODS Cell nuclei were prepared from SMMC LTNM hepatoma which is a transplanted human liver cancer born on nude mice. Samples were irradiated with 60 Co γ rays at different doses or dose rates. N ethylmaleimide (NEM) and ddTTP were used as selective inhibitors to DNA polymerases. The reaction of DNA repair synthesis was carried out with the selective inhibitor test. RESULTS It was found that the 3H TTP incorporation in irradiated nuclei or calf thymus DNA was significantly higher than that in the non irradiated ones, under the conditions of DNA polymerase α or γ being inhibited. When NEM and ddTTP which selectively inhibits DNA polymerase β both existed in the DNA repair synthesis reaction mixture, the 3H TTP incorporation in irradiated DNA did not significantly increased. Furthermore, 3H TTP incorporation into DNA of SMMC LTNM hepatoma nuclei was higher than that of normal hepatocyte nuclei ( P <0 01). The DNA repair synthesis induced by DNA polymerase β reacted more fast in hepatoma nuclei than in hepatocyte nuclei. CONCLUSION The effects of DNA repair synthesis induced by DNA polymerase β in some tumor cells might be stronger than that in normal cells, which may facilitate the cells to repair DNA damages from radiation.

Public Screening for Early Carcinoma of Gastric Cardia: Rule of Carcinogenetic Development Observed by Endoscopy

Objective： To study the rule of development of early cancer of gastric cardia in vivo in public screening. Methods： A prospective cohort study on gastric cardiac cancer was performed in the high incidence area of cancer of esophagus and stomach in china. 106 subjects had been examined regularly by endoscopy to observe the change of mucosa in high incident area of gastric cardiac carcinoma developing at the root of gastric cardiac ridge by taking biopsy specimen. All specimens were diagnosed through normal pathological process to study the prognosis of pro-cancer lesions of gastric cardia. Results： The results of 106 subjects who had been observed for 4 years were： （1） Of 8 normal persons, 3 stayed normal, 4 turned to chronic gastritis, 1 developed early gastric cardiac cancer. （2） Of 61 persons with chronic gastritis, 11 were observed to have gland atrophy, 4 mild atypical hyperplasia, and 2 highly atypical hyperplasia. （3） Of 9 subjects showing atrophic chronic gastritis, 5 revealed no change, and 4 became chronic gastritis. （4） Of 22 subjects who revealed mild atypical hyperplasia, 17 resolved, 4 showed no change, and 1 advanced to highly atypical hyperplasia. （5） One person with highly atypical hyperplasia reverted to mild atypical hyperplasia. （6） Of 5 subjects with early gastric cardiac cancer without any treatment, 1 became advanced cancer, 1 still stayed in early cancer stage, and 3 turned to atypical hyperplasia. Conclusion： The development of early cancer of gastric cardia would proceed through the stages of chronic gastritis, gland atrophy, and atypical hyperplasia. （2） The early cancer and pre-cancer lesions of gastric cardia is reversible, though possessing malignant possibility.

Intermittent portal triad clamping in resection of liver tumors involving the hepatocaval confluence

Objective: To review our experience in and the re-sults of resecting liver tumors involving the hepatoca-val confluence under intermittent portal triad clam-ping (PTC).Methods: Sixty-eight consecutive patients with livertumors involving the hepatocaval confluence under-went hepatectomies with liver parenchymal transec-tions under intermittent PTC.Results: All the tumors were successfully resected un-der PTC, except for one in which the infrahepatic in-ferior vena cava was concomitantly occluded in addi-tion to PTC. There was neither operative death noruncontrollable massive bleeding or air embolism oc-curred in our patients. The bleedings from the mainand short hepatic veins and right adrenal veins wereproperly managed during the operation, with a meanintraoperative blood loss of 1400 ml. Of the 68tumors resected, 65 were hepatocellular carcinomas(HCC). Their 1-, 2-, 3- and 4-year suvival rateswere 64.11%, 52. 82%, 44.90% and 36.98%, re-spectively, and the patients with HCC with capsulessurvived significantly longer than those with HCCwithout capsules.Conclusions: The liver tumors involving the hepato-caval confluence could be safely resected simply un-der PTC, without routine use of total hepatic vascu-lar exclusion. As for HCCs in this area, the tumorwith capsule is a better indicator for surgical resec-tion than that without capsule.

INVESTIGATION OF THE RELATIONSHIP BETWEEN CELL PROLIFERATIVE ACTIVITY AND INVASION, METASTASIS AND PROGNOSIS OF GASTRIC CANCER

To evaluated Bromodeoxyuridine (BrdUrd) /DNA doubleparametric method in detection of gastric carcinoma and to analyze the relationships of cellular BrdUrd labeling indices(LI), G2/M-phase fraction(G2/MPE) and DNA content to the depth of invasion, lymphatic vessel invasion, lymphatic node metastasis, peritoneal dissemination and blood vessel invasion and prognosis. Methods: Fresh tumor samples from 60 cases were examined by BrdUrd/DNA doubleparametric flowcytometry. Propidium iodide(PI) was used as fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used as second antibody. Results: The values of BrdUrd LI in patients with serosa invasion was significantly higher than those without serosa invasion (P<0.01); there was statistical significance in 5-year survival rate between the two groups (P<0.01). Both BrdUrd LI and G2/MPF values were significantly higher in patients with lymphatic vessel invasion than those without invasion (P<0.01); the patients with lymphatic vascular invasion carried a significantly poor prognosis (P<0.01). Both BrdUrd LI and G2/MPF values were significantly higher in patients with lymphatic node metastasis than those without metastasis (P<0.01), there was a statistical significance in 5 years survival between these 2 groups. The incidence of lymphatic node metastasis was significantly higher in aneuploid carcinoma (P<0.05), and the patients with aneuploid carried a significantly poor prognosis (P<0.05). Patients with peritoneal dissemination had a significantly worse prognosis (P<0.01). G2/MPF values were significantly higher in patients with blood vessel invasion than those without invasion (P<0.01). Conclusion: Cellular BrdUrd LI, G2/MPF and DNA content are related to depth of invasion, lymphatic vessel invasion, peritoneal dissemination, blood vessel invasion and prognosis of gastric carcinoma.

Evaluate the Expression of uPA,PAI-1 in Human Gastric Cancer and its Correlation with the Angiogenesis by the Application of Tissue Microarray

OBJECTIVE To investigate the expression of urokinase-type plasminogen （uPA）, its inhibitor-1 （PAI-1） mRNA and its protein in human gastric cancer and to find out the relationship among the tumor differentiation, angiogenesis, and other clinical pathologic factors. METHODS In situ hybridization （ISH） was used to get the uPA, PAI-lmRNA in 110 cases with human gastric cancer in 2-tissue microarray （TMA）. Immunohistochemical staining （S-P method） for uPA, PAI-1 protein and CD34 were performed in the 110 cases in 2 TMA. RESULTS The expression of the uPA, PAI-lmRNA and their protein happened in the cytoplasm of gastric cancer cells were induced by the poor differentiation of the GC, and the expression of uPA had an increasing trend while the expression of the PAI-1 had a decreasing trend. The microvessel density （MVD） had a positive correlation with the clinical stages and the significant relationship with the lymph node metastasis （P 〈 0.05）. The MVD in uPA positive group was significantly higher than those in uPA negative group （P 〈 0.05）. The expression of PAI-1 has no correlation neither with the clinical stages nor the lymph node metastasis. CONCLUSION The uPA play an important role in invasion and metastasis of GC through promoting angiogenesis. Interdicting the secretion and function of the uPA may allow the target therapy against the tumor invasion. As a new high-throughput technology, the tissue microarray is a valuable way to be used in clinical treatment.

Expression of galectin-3 correlates with apoptosis in pituitary adenoma cells

Objective To investigate the expressions of Galectin-3 （Gal-3）, Bcl-2 and Bax in human pituitary adenomas, and to explore the interrelation among them. Methods RT-PCR and immunohistochemistry were applied to detect the mRNA and protein expressions of Gal-3, Bcl-2 and Bax in surgically excised human pituitary adenoma tissues, including invasive and non-invasive pituitary adenomas, and the correlation analysis was performed. Results The Gal-3 and Bcl-2 expressions in the invasive pituitary group were significantly higher than those in the non-invasive group, and the expression of Bax had no significant difference between the two groups. Pearson＇s correlation analyses showed that the Gal-3 expression was posi- tively correlated with Bcl-2, but was not correlated with Bax, which was inversely correlated with expression of Bcl-2. Conclusion Gal-3 may function through a cell death inhibition pathway involving Bcl-2 to enhance cell proliferation, which result in the invasive growth of pituitary adenoma. These results indicate that Gal-3 has an important role in pituitary tumor cell proliferation and may serves as a possible therapeutic target in treatment of pituitary tumors.

Clinical Study of Multi-drug Resistance Gene(MDR1) Expression in Primary Ovarian Cancer

This study was designed to measure the multi-drug resistance gene (MDR1) mRNA content and analyze clinical relationship between MDR1 expression and drug resistance in primary ovarian cancer. Reverse transcription PCR (RT-PCR) was used to measure MDR1 mRNA content in biopsy sample of 31 primary ovarian cancers (experimental group) and 30 gynecological tumors (control group). The level of 95.2% (20/21) MDR1 expression was relatively low, and the detected rate of MDR1 expression was 67.7%(21/31) in experimental group,which was higher than that in control group (40.0%, P<0.05). The differences of MDR1 expression between the effective group and no effect group after combined chemotherapy was significant (P<0.05). No significant relationship was found between MDR1 expression and clinical stage or histological classification or grade of differentiation in experimental group. We are led to concluded that primary ovarian cancers have drug-resistance clones which might express MDR1 spontaneously and expression of MDR1 may be used as a prognostic and predictive indicator for clinical response of ovarian cancers to combined chemotherapy.

Effects of arsenic trioxide on drug transporting molecules in multidrug resistance malignant neoplasma MR2 cell line

Objective： To study the effect of arsenic trioxide （As203） on the expression of drug transporting molecules in multidrug resistance malignant neoplasma acute promyelocytic leukemia （APL） MR2 cell line. Methods： MR2 resistant to alltrans retinoic acid （ATRA） and non-ATRA resistant APL cell line NB4 were used. Expressions of P-glycoprotein （Pgp）, multidrug resistance protein （MRP） and lung resistance-related protein （LRP） were detected by immunocytochemical assay. Results： The expression of Pgp was significantly higher in MR2（30%-40%） than that in NB4（10%-20%） （P 〈 0.001）, and the expression of MRP was also higher in MR2 （56.9 ± 3.4 - 21.2 ± 1.1） than that in NB4 （20.6 ± 5.3 - 16.7 ± 1.2） （P 〈 0.001）. As2O3 ranging from 0.5-2.0 μmol/L, could significantly decrease the expressions of Pgp and MRP. The expression of Pgp and MRP in MR.2 cell line were negatively correlated with the dose and duration of action of As2O3. Conclusion： Pgp and MRP may be the sensitive targets of As2O3 to overcome drug-resistance. ATRA might be the substrates of Pgp and MRP.

Phosphorylated KDR can be located in the nucleus of neoplastic cells

KDR （kinase insert domain receptor） phosphorylation induces several effects which lead eventually to cell proliferation and survival. The precise mechanisms by which KDR, once it is activated, communicates with the nucleus are starting to be understood but have not yet been completely unravelled. Two in vitro studies on animal cell lines reported in the literature have demonstrated that, following stimulation with VEGF, KDR is actually translocated within the nucleus. Our aim was to investigate whether this translocation occurs in human cells both in vitro and in vivo. Using laser scanning confocal microscopy, a variable nuclear localization of phosphorylated and total KDR in cell lines and tumour samples was found. In human neoplastic cell lines, hypoxic stimulation greatly increased the nuclear amount of total KDR but less so that of the phosphorylated form. Only after hypoxia and VEGF stimulation there was a comparably increased expression of phosphorylated and total KDR observed in the nuclei of these cells. We conclude that neoplastic cells show a variable expression of total and phosphorylated KDR in the nucleus. The precise functional meaning of nuclear location remains to be established.

Reversal Efficacy of SDZ-PSC833 and Verapamil on Drug-Resistant Human Cervical Carcinoma Cells Line

This article discusses the characteristics of the drugresistant human cervical carcinoma cell line HeLa/MMC and investigates reversal effect of SDZ-PSC833（PSC833） and Verapamil （Ver） upon HeLa/MMC. Relative content of cell DNA detection, H3-TdR incorporation test and drug-resistance have been experimented, meanwhile a tumor transplant model in the nude mice for Hela/MMC has been established. Then the volume of cancer, tumor weight, curve line of growing up in tumor -bearing nude mouse and its morphological variation of carcinoma organism are observed to evaluate the reversal effect of PSC833 and VER. It has been found that： （1） Compared with HeLa cells,subline HeLaJ MMC Cells＇ drug-resistant capability against Mitomycin（MMC） or cisplatin（DDP） is as 5.02 folds or 2 folds as that of the former, respectively. The Hela/MMC cell line has characteristics of multi- drug resistance and stronger multiplication; （2） PSC833 and VER can reverse resistance of HeLa/MMC to Mitomycin in vivo.PSC833 will be a better candidate for reversing multidrug resistant than verapamil in clinic application.

INFLUENCE OF ALPHA-FETOPROTEIN ON THE GROWTH OF TUMOR CELLS IN VITRO

It has been recognized that alpha-fetoprotein (AFP),as an oncofetal antigen, re-expresses in large amounts inadult tumor cells and serves clinically useful purposes asa tumor marker assay. However, its biological activiticsare still far from clear. In thc present study, the ability ofAFP to stimulate tumor cell growth was observed by invitro test system. The new finding indicates that AFPcontributes to the generation and development of tumorand is an important target action site of tumor therapy.

PROGNOSTIC FACTORS ANALYSIS FOR STAGEⅠ RECTAL CANCER

Objective: To explore the death-related factors of stageⅠrectal cancer patients. Methods: 89 cases of stage I rectal cancer patients between 1985 and 2000 were retrospectively studied for prognostic factors. Factors including age, gender, tumor size, circumferential occupation, gross type, pathological type, depth of tumor invasion, surgical procedure, adjuvant chemotherapy and postoperative complication were chosen for cox multivariate analysis (forward procedure) using Spss software (10.0 version). Results: multivariate analysis demonstrated that muscular invasion was an independent negative prognostic factor for stageⅠrectal cancer patients (P=0.003). Conclusion: Muscular invasion is a negative prognostic factor for stage I rectal cancer patients.

ANTITUMOR EFFECT OF INTRATUMORAL INJECTION OF LIPOSOME-ENCAPSULATED G-CSF GENE AND IN SITU BIOLOGICAL CHARACTERISTICS OF THE TREATED TUMOR CELLS

This word was supported by grant from Military Medical Research Foundation of china (96z032). ** To whom correspondence and requests for reprints should be addressed. This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 1997). In order to investigate the antitumor effects of the in vivo G CSF gene therapy mediated by liposome and its mechanisms, human G CSF gene was encapsulated into liposome and was directly injected into tumor mass of C 26 colon adenocarcinoma bearing mice. After direct intratumoral injection of liposome encapsulated G CSF DNA, the subcutaneous tumor growth was dramatically inhibited and the survival time was prolonged signifi cantly. Tumor regression could be observed in about 30% of C 26 bearing mice. By the analysis of the antitumor mechanisms, we found that anti G 418 (600ug/ml) clone could be selected from the tumor cells freshly separated from the treated C 26 tumor mass, and secretion of G CSF in the supernatant could be detected. Northern blot also confirmed the expression of hG CSF by the tumor cells. Higher expressions of MHC class I(H 2k d) molecule and ICAM 1 on the tumor cells could be observed. The results demonstrated that liposome can effectively transfect G CSF gene into tumor cells in situ , and then increase the immunogenicity of the tumor cells which may contribute to the activation of the local antitumor immune responses effectively.

Expressions of Connexin and Par-3 in the Distal Margin of Rectal Cancer after Ultra-low Anterior Resection

This study examined the expression of connexin and protease-activated receptor 3 （par-3） in the distal resection margin of rectal cancer and the correlation of the expression of the two proteins with tumor relapse. A total of 40 patients with rectal cancer underwent ultra-low anterior resection with curved cutter stapler. The pathological specimens were divided into 3 groups in terms of sampling sites： tumor group, 2.0-cm group （in which the tissues were harvested 2.0 cm distal to the tumor tissues）, 3.0-cm group （in which the tissues were taken 3.0 cm away from the tumor tissues）. All the samples were pathologically observed and then measured for the expression of connexin and par-3 by employing immunohistochemistry and Western blotting. The operations in this series went uneventfully. No anastomotic stoma bleeding, stenosis and death occurred postoperatively. Histopathologically, in the tumor group, epithelial cells lost normal pattern of arrangement and polarity, and were loosely connected and even detached. In the 3.0-cm group, the epithelia had normal appearance, obvious cell polarity and essentially intact cell junction. Immunohistochemistry and Western blotting indicated that the 3.0-cm group had the strongest expression of connexin and par-3, and the expression in the 2.0-cm group and the tumor group was relatively weak. There existed significant difference in the expression of the two proteins among the three groups （P〈0.05 for all）. It was concluded that the down-regulated connexin and par-3 in the distal margin of rectal cancer tissues may indicate the progression of the disease and high likelihood of recurrence and metastasis. Although no tumor cells were found in the sections of the 2.0cm group, the decreased expression of connexin and par-3 may suggest the development of anaplasia and the increased odds of tumor relapse. Therefore, we are led to speculate that tumor resection only including 2.0 cm of unaffected rectum could not completely avoid the distant metastasis and local relapse.

ESTABLISHMENT OF CRITERIA FOR MEASURING MDR-1 GENE EXPRESSION LEVEL IN BREAST CANCER BY RT-PCR

Objective: To formulate criteria of multidrug resistance (mdr1) gene expression for predicting chemotherapy response and prognosis. Methods: Using reverse transcriptionpolymerase chain reaction (RTPCR) assay, the expression of mdr1 gene in 82 breast cancer samples were detected. Results: The data were treated by statistic analysis system (SAS)singlevariate analysis. It showed that the level of mdr1 gene expression clearly deviated from normal to right distribution (P<0.0001), and thus might be divided by quantiles P50 (mdr1/β 2MG=0.2) and P75 (mdr1/β 2MG=0.6), which were taken as the preliminary criteria for analyzing 56 patients' chemosensitivity to ADM、VDS and VCR in vitro and 32 relapsed metastatic patients' chemotherapy response in vivo, seperately. When mdr1/( 2MG(0.2, the ratios of resistance gradually escalated, but there were about 30%～50% of the cases who showed sensitive to the drugs in vitro and effective to chemotherapy in vivo. When mdr1/β 2MG≥0.6, the most of patients showed drug resistance both in vitro and in vivo. Conclusion: According to the abovementioned results, criteria of evaluating mdr1 gene expression level was formulated: the mdr1/β 2MG<0.2 (P50) was considered as negative expression, the ratio≥02～<0.6 (P75) was weakly positive expression, ≥0.6 was strongly positive expression. This indicated that different levels of mdr1 gene expression may reflect objectively drug resistance in vitro and chemotherapy response in vivo.

THE CLINICAL EVALUATION OF ANGIOGENESIS INHIBITOR-20(R)-GINSENOSIDE RG_3 IN LUNG CANCER

Objective The aim of the study was to make a further evaluation of Ginsenoside Rg3. Methods The clinical effects of the drug on moderate and advanced lung cancer, including side effects, were observed. Results Ginsenoside Rg3 improved chemotherapy significantly. The clinical relief rate of patients treated with antiangiogenic agent 20 (R) Ginsenoside Rg3 was 36.6%, which was higher than that of the patients not treated with it (16.7%)( P <0.05). It had no significantly different effects on lung cancers of different types of tissues ( P >0.05). It provided better treatment on the cancer at early stage than that at advanced stage ( P <0.05). Moreover the living qualities of the patients were improved notably ( P <0.05). Conclusion Combined with chemotherapy, angiogenesis inhibitor 20(R) Ginsenoside Rg3 can improve clinical therapeutic efficacy and the living qualities of patients significantly.

Cancer Prevention through Legislation Hong Kong Experience

Cancer is the major cause of death worldwide and in the local community. Due to the aging population and changes in lifestyle of the citizens, it is expected that the incidence of cancer will continue to increase. In fact, according to the World Health Organization, about 30% of cancer death can be prevented. The fight against cancer relies on support from the government, together with collaborations with the policymakers, healthcare professionals, and the public. Legislation can act as a tool for cancer prevention. The purpose of this paper is to provide an overview of the global cancer burden and to describe how cancer legislation acts as a tool for cancer prevention in the Hong Kong region.