Study On The Intraperitoneal Free Cells During Operation And Its Related Gene In Patients With Advanced Gastric Cancer

Objective: To research the positive rate of the peritoneal lavage fluid positive rate of exfoliated cells and DNA aneuploid,which from Operation process before and after of the patients with advanced gastric cancer,and to evaluate the relations of carcinoembryonic antigen(CEA),cytokeratin- 7(CK7),matrix metalloproteinase 7(MMP7) and other related gene,clinical and pathological and the effect of operation.Methods:Select a prospective,open,randomized,multi center phase III clinical trials in patients with advanced gastric cancer(NCT01516944) in 132 cases of direct operation,all the patients underwent total abdominal plain and enhanced CT imaging to detect distant metastasis,and after the gastroscope bite seized 6~8 pieces of tissue,pathological diagnosis of gastric adenocar- cinoma,and do not use chemotherapy or radiotherapy.Radical operation were performed in standard D2,if there is metastasis when the operation opened,it is suitable conditions for palliative operation,the whole process of operation with no tumor operation strictly.After the beginning of surgery in patients with abdominal exploration,with position of the first high pin low,lift the transverse mesocolon,use saline with 300~400ml(37℃) to flush the abdomen and the Doglas fossa,use the negative pressure aspirator directly suction the lavage fluid into the clean specimen bag,if ascites is directly extracted,collect about 300 ml,this is the former group specimens of the surgical procedure;at the end of surgery using sterile distilled water rinse 3 times before abdominal closure(37℃) about 10 minutes,suction clearly,and use normal saline 300~400ml(37℃) flushing all the abdominal cavity,use a new negative pressure aspirator suction aspiration lavage fluid about 300 ml.This is the process of specimens after surgery group two specimens were performed,cytology of peritoneal lavage fluid(peritoneal lavage cytology,PLC) examination,and the use of Feulgon staining to detect the DNA ploidy,ploidy abnormalities are more than 5c,the four chart report,the first picture of the horizontal axis is the content of DNA,the vertical axis is the number of cells,the normal value is less than 5c.The second chart x-axis DNA content,the longitudinal axis of the nuclear area.The third picture for the first 20 DNA the highest DNA content ploidy value and image.Finally,a DNA ploidy for numerical maximum endoscopic image.Results are reported for the(negative) no DNA ploidy abnormal cells;(positive) visible DNA ploidy abnormal cells.Simultaneous detection of CEA,CK7,MMP7,observe the target cells in the slides(considered as cancer cells and except to inflammatory cells,tissue cells and hyperplastic mesothelial cells).CEA and MMP7 located in cytoplasm,cytoplasmic uniform like with brown determined as positive,no stain is negative;CK7 localized in the cytoplasm,coloring determined as positive,otherwise negative;all specimens with WT-1,Calretinin to exclude mesothelial cells.Analysis of the relationship of PLC and DNA aneuploid was positive and the clinicopathological para- meters and effect of operation.Results:1 At present,PLC is considered to be the detection of exfoliated cancer cells(exfoliated cancer cells,ECC) of the gold standard,the operation method is simple,but its low sensitivity.2 In 132 cases of patients with gastric cancer Operation process before group PLC was positive in 24 cases(18.18%),while aneuploid positive in 35 cases(26.52%),with significant difference(P=0.001),the sensitivity was 100%, specificity was 89.81%.3 The positive rate of the Operation process before group of CEA,CK7 and MMP7 were 27.27%,24.24% and 25%;and the positive rate of the after operation CEA,CK7, MMP7 were 15.15%,14.39%,12.88%.The positive rate of PLC positive and CEA, CK7 differences(P<0.05),but the positive rate of DNA of different ploidy positive and CEA,CK7,MMP7 no significant differ- rence(P>0.05).4 The positive rate of radical operation of R0 PLC was 12.07%,signific- antly lower than the R1 or R2 radical or palliative operation 62.5%(P<0.05).5 Operation process before group peritoneal lavage fluid positive and the tumor diameter,depth of invasion,lymph node metastasis,distant metastasis, TNM stage,surgical cure of related(P<0.05),has nothing to do with gender,age,tumor location,differentiation degree,Lauren type,Borrmann type(P>0.05).6 Full use of strict operation without tumor after operation,a large number of closing the abdomen before rinsing with distilled water,the positive rate of PLC after operation process group was 9.09%,compared with the preoperative 18.18% has decreased significantly,and there are significant differences between them(P=0.004).Surgery patients Operation process before group PLC positive in 24 cases,14 cases became negative after operation process;however,there are two patients in Operation process before group PLC was negative(visible atypical cells,and DNA ploidy were greater than 5c),and the procedure after the change to positive,the 2 patients in Operation process before group with the results of PLC may be false negative,and the posto- perative results as true positive;also may be iatrogenic tumor cells caused by exfoliative of improper operation.Conclusions:1 PLC method is still the gold standard,but its sensitivity is low,the positive rate is high,and the positive rate of detection results of DNA ploidy and CEA,CK7,MMP7 and other related genes is higher than PLC.2Operation cure degree is higher,the positive rate of PLC is low.3 This is also related with tumor staging,with the depth of tumor invasion,lymph node metastasis,distant metastasis,TNM stage increased,the radical operation success rate is low,the positive rate of PLC will increase.4 In the process of operation by no tumor procedure strictly,can significantly reduce the cancer cells exfoliated planting.