The Study Of T-cell Subgroups And Tungstens As The Influential Factor Of Gastric Carcinoma Relapse And Metastatic And The Correlation Of T-cell Subgroup And Tungstens

Objective:In order to study the influential factors of relapse and metastatic in gastric carcinoma from different aspects, we detected tungstens including copper, zinc, iron, calcium, magnesium etc. and T-cell subgroups including CD3~+, CD4~+, CD8~+, CD4~+/CD8~+ etc. And to investigate the age, gender, staging, course and family history. Meanwhile we analyzed if there were noticeable differences the T-cell subgroups, tungstens in normal population and in carcinoma patients, and discussed if T-cell subgroups and tungstens had correlation or not, then found the inherent connections between tungstens and immune function.Material:Selected 60 gastric cancer patients as study object at random, including 32 males and 28 females, the meta-age was 67 years old. All of the patients, there were 36 patients who had relapsed and metastasized and 24 patitents who had not. All patients had final diagnosis by biopsy or postoperative pathology. According to the TNM stage methods of American Carcinoma Joint Committee(AJCC), 8 patients were in theⅠstage, 28 patients were in theⅡstage, 20 patients were in theⅢstage, 4 patients were in theⅣstage. The cases who had relapsed and metastasized were all certified by pathology or CT methods. Their blood preparation were under routine examination at pretherapy. Before, the patients those who were to inter the group were not be treated by chemo, radiotherapy and immunize within 3 weeks. As well as to remove the other correlated disease. To obtain empty stomach vein blood of the patients, about 4ml and placed them into two test tubes, one was 1ml, no anticoagulation, centrifugated, separated serum, enclosed, then to preserve in the refrigerator which is -20℃, stayed to measure tungstens. Another was 3ml, anticoagulated with 2% heparin, determined the nucleolus/nucleus of lymphocyte, T-cell subgroups, NK cell.Methods:Determination of T-cell subgroups and NK cellUsing SAP detection kit and the method of SAP. Separated mononuclear cell with lymphocytes separating medium sec- undum artem to prepare cell smear, standby after stoving. Prepared color developing reagent to standby. Drawed a circle outside the example on the thorough desiccated cell smear, immobilization, admoved appropriate amount antihuman CD3~+,CD4~+,CD8~+,CD57 IL-1ra into hygro-casing, stayed overnight under 4℃, eluted 3 times with PBS. Droped appropriate amount biotin labeling anti-mice Ig-G, brooded them in 37℃, eluted 3 times with PBS. Droped appropriate amount alkaline phosphatase labelling strepto-antibiotin, brooded in 37℃, eluted 3 times with PBS. Droped chromogenic agent, made it colorated in room temperature. To observe under the microscope, eluted with PBS when the cell membrane appeared red markers. Droped cell nucleus counter staining solution, eluted it with tap water after 30 seconds. Droped mounting agent, cover plate and mount, tested under microscope. Counted 300 mononuclearcell in well staining visual fields under microscope which had red markers on the surface was masccline cell.Worked out the masccline rate.The detection of nucleolus/ nucleiObtained 1ml venous blood and influxed into culture flask that contained culture solution, then infunded plastics test tube and centrifugate. To throw away the supernatant, added hypotonic liquor, misced bene. Placed it in the thermostatic waterbath casing which was 37℃halfhour. Misced bene with stationary liquid. Centrifugated, threw away the supernatant, misced bene with 5ml stationary liquid and centrifugate. To throw away the supernatant and to stay suspension, droped onto the glass, airdryed. First added 2 drops A liquor onto the glass, then added 1 drop B liquor, placed the cover glass, put it into the baker until there were mass golden yellow gas bubbles appeared. Washed it cleanly.Trace elementAdopted the method of atomic absorption spectroscopical photometer. Various kinds of elemental hollow cathode lamp. Prepared different level standard liquor with the simplex standard solution, and drawed the standard curve. The tentative water was second order deion water. The serum specimen was digestion processed by normal pressure wet method and yield the data with the machine.Statistical analysisHandled the data with the SAS 9.0 statistics software, all groups of the data were paragraphed and evaluated, and imported the data into computer by means of x or y. Firstly used simplex factor logistic regression analysis to reject the nonsense factors according to the norm ofα=0.1. The other factors were analysised by the multiple factor logistic stepwise regression. Compared the means of every group and the means of normal controls with T-test. Firstly used test of normality to analysis the T-cell subgroups and tungstens, the data that correspond the test were to use linear correlation analysis, and the data that discrepancy were to use rand correlation analysis. According to the level ofα=0.05,Ρ<0.05 meant that the difference was significant,Ρ>0.05 meant that the difference was not significant.Results:1 There are 36 patients who have relapsed and metastasized in the 60 patients of gastric carcinoma, account 60% (36/60).2 The result of the simplex factors logistic regression ana- lysis shows that, clinic patho-TNM staging (x=2.0526, P=0.0001, OR=7.788), the metastasize of lymph node (x= 0.9069, P=0.0932, OR=2.477), the course of disease (x=1.4826, P=0.0016, OR=4.405), the masculine cell population of CD3~+(x=-1.3753, P=0.0521, OR=0.253), the masculine cell population of CD4~+(x=-1.5804, P=0.0057, OR=0.206), the masculine cell population of CD8~+(x=-0.7002, P=0.0615, OR=0.496), the masculine cell population of NK(x=-0.7002, P=0.0615, OR=0.496), the contents of Cu(x=4.1528, P=0.0002, OR=6.61), the contents of Mn(x=1.6573, P=0.0088, OR=5.245), Cu/Zn(x=2.1972, P=0.0003, OR=9.0), the ten index are related with the relapse and metastasize of gastric carcinoma (Ρ<0.1). Besides the factors above, the simplex factors logistic regression analysis of sex, age, family history, nucleolus/nuclei of T-cell,CD4~+/CD8~+ and Zn, Fe, Ca, Mg have not statistically significant(Ρ>0.1).3 The result of multiple factor logistic correlation analysis shows that the clinic patho-TNM staging(x=3.2046, P=0.0074, OR=24.645), and the contents of Cu(x=4.1528, P=0.0002, OR=63.61)are related with the relapse and metastasize of gastric carcinoma(Ρ<0.1).4 The nucleolus/nuclei, the masculine cell population of CD3~+, CD4~+, CD4~+/CD8~+ are lower than the control group obviously, the masculine cell population of CD8~+ is top than the control group(P<0.01). The masculine cell population of NK is lower than the control group(P<0.01).5 The contents of Cu,Zn,Fe,Mg of the gastric cancer patients are all lower than normo-control group, the contents of Ca, Mn and the ratio of Cu and Zn are top than the normal control group(P<0.01).6 There is positive correlation between the nucleolus/ nuclei and Fe, Mg(P<0.05), negative correlation between CD3~+ and Fe, Mn(P<0.05), positive correlation between CD4~+ and Zn, Ca(P<0.05), negative correlation between CD4~+ and Mn(P<0. 05), negative correlation between CD8~+ and Ca(P<0.05), positive correlation between CD4~+/CD8~+ and Zn, Ca(P<0.05), negative correlation between CD4~+/CD8~+ and Cu/Zn(P<0.05), negative correlation between NK and Mn(P<0.05).Conclusions:1 The rate of relapse and metastasize of gastric cancer is high.2 The metastasize of lymphonode, clinic patho-staging, the length of course of disease, CD3~+, CD4~+, CD8~+, NK, Cu, Mn, Cu/Zn are all the influential factors of relapse and metastasize of gastric cancer. The broader the metastasize of lymphonode is, the later the clinic patho-staging is,the longer the course of disease is, the fewer the masculine cell population of CD3~+, CD4~+, CD8~+, NK is, the higher the contents of Cu, Mn, Cu/Zn is, the possible the risk of gastric cancer of relapse and metastasize is. These risk factors could be used as significant indexes for the prediction of relapse and metastasize of gastric cancer.3 The immune function of gastric cancer patients is lower than that of the normal control obviously. The level of Cu, Zn, Fe, Ca, Mg, Mn in the blood of the gastric cancer patients is lower than that in the normal control obviously, and the contents of Ca and Mn are higher than that in the normal control, which can offer evidence for clinical application of immunotherapy and elementum therapy.4 There are correlations between some tungstens and the immune function. It hint that we can give elements to the patients of gastric cancer or to avoid the intake of destructive elements, thereby to raise the disease resistance of organism and restrain the relapse and metastasize.