To evaluate the change of erythrocytes malondialdehyde in patients with colon cancer before and after radical surgery and find out the relationship between the changes of erythrocyte malondialdehyde in peripheral veins after surgery and some surgical characteristics. Subjects and methods: 60 patients with color cancer treated with radical surgery from March 2015 to January 2017 at 103 Military Hospital were included in the research. The characteristics of the patients and the resection of the colon tumor were recorded, the oxidative stress was determined by measurement of erythrocyte malondialdehyde, blood samples were collected from the peripheral veins at 4 times: before surgery, 1 day after surgery, 3 days after surgery and 7 days after surgery. Results: 1 day after surgery, erythrocyte malondialdehyde increased compared to preoperation (p < 0.05). 3 days and day 7 after surgery, erythrocyte malondialdehyde decreased compared to 1 day after surgery but compared to preoperation, the change was not significant with p > 0.05. Conclusion: Compared to preoperation, the peripheral erythrocyte malondialdehyde increased significantly at the point of 1 day after surgery, and over time the erythrocyte malondialdehyde index tended to decrease.
Journal of military pharmaco-medicine n02-2019 INVESTIGATION ON CHANGES IN THE ERYTHROCYTE MALONDIALDEHYDE VALUE IN PATIENTS WITH COLON CANCER AFTER RADICAL SURGERY Pham Manh Cuong1; Nguyen Van Xuyen1; Trinh Hong Thai2 SUMMARY Objectives: To evaluate the change of erythrocytes malondialdehyde in patients with colon cancer before and after radical surgery and find out the relationship between the changes of erythrocyte malondialdehyde in peripheral veins after surgery and some surgical characteristics Subjects and methods: 60 patients with color cancer treated with radical surgery from March 2015 to January 2017 at 103 Military Hospital were included in the research The characteristics of the patients and the resection of the colon tumor were recorded, the oxidative stress was determined by measurement of erythrocyte malondialdehyde, blood samples were collected from the peripheral veins at times: before surgery, day after surgery, days after surgery and days after surgery Results: day after surgery, erythrocyte malondialdehyde increased compared to preoperation (p < 0.05) days and day after surgery, erythrocyte malondialdehyde decreased compared to day after surgery but compared to preoperation, the change was not significant with p > 0.05 Conclusion: Compared to preoperation, the peripheral erythrocyte malondialdehyde increased significantly at the point of day after surgery, and over time the erythrocyte malondialdehyde index tended to decrease * Keywords: Colon cancer; Oxidative stress; Erythrocyte malondialdehyde INTRODUCTION Colon cancer is one of the most common cancers with high morbidity and mortality rates in many countries around the world In Vietnam in 2010, along with rectal cancer, colon cancer ranked fourth after lung, stomach and liver cancer in men, ranked fifth after breast, stomach, lung and cervical cancers in women [1] In the body, there is always a balance between the production of oxidants and their elimination with antioxidants, when this process is imbalanced by free radicals that exceed the antioxidant capacity, may lead to oxidative stress Consequently, biological molecules such as DNA, proteins, lipids are constantly being attacked by free radicals that cause defects in biological molecules, damage to cells and tissues, and may in particular cause alteration gene Oxidative stress has also been identified in relation to the initiation and progression of many different cancers, including colon cancer [2, 3] 103 Military Hospital Hanoi University of Science, Vietnam National University Corresponding author: Pham Manh Cuong (famcuong103@gmail.com) Date received: 20/12/2018 Date accepted: 23/01/2019 213 Journal of military pharmaco-medicine n02-2019 Radical surgery is still the main method for treating colon cancer, but the trauma in surgery also causes oxidative stress for the body due to the release of many free radicals The research on changes in postoperative oxidative stress has also been investigated in many different surgeries and in surgical treatment of colon cancer [4, 5] In Vietnam, the assessment of oxidative stress has been carried out on many different diseases, but no studies have been done for colon cancer alone Assessing the status of oxidative stress through the determination of malondialdehyde (MDA) in blood, a product of lipid peroxidation, is widely and popularly applied in many studies Therefore, the aim of this research is: To evaluate the change of erythrocyte MDA in peripheral vein in patients with colon cancer after radical surgery, to find out the relationship between the changes of erythrocyte MDA after surgery according to a number of surgical characteristics SUBJECTS AND METHODS Subjects 60 colon cancer patients were treated by radical surgery at Department of Abdominal Surgery, 103 Military Hospital from March 2015 to January 2017 * Inclusive criteria: Results of postoperative pathology as adenocarcinoma of colon, conducted radical surgery (R0) in accordance with the prescribed procedure and has a full quantitative result of erythrocyte MDA before and after surgery * Exclusive criteria: Body mass index (BMI) ≥ 30 kg/m2, smoking, alcoholism, 214 systemic inflammatory disease, diabetes, cardiovascular disease, autoimmune diseases, patients with complications such as obstruction, perforation colon or distant metastasis Method * Research design: A cross-descriptive research * Research content: The research group was divided into small groups, including the following criteria: age (< 60; ≥ 60); gender (male; female); disease stage (according to TNM I; II; III), surgical method (laparoscopic surgery; open surgery), surgical level (expanded; not expanded) The extent of surgery that has been expanded is defined as being in addition to colectomy, the patients were performed other combined surgeries such as gastrectomy, resection of small intestine, oophorectomy, etc or must proceed to total colectomy Indicators of oxidative stress were determined by quantifying erythrocyte MDA at times: before surgery, day, days and days after surgery * Surgical procedure: Before surgery, all patients were prepared to clean the colon and taken the Fleet Phospho Soda manufactured by C.B Fleet Company Inc (USA) in the morning before surgery All patients were induced with propofol (3 - mg/kg), fentanyl (1.51 g/kg) After endotracheal intubation, the patients were provided with mechanical ventilation using an air mixture containing 30% O and maintained anesthesia with sevoflurane (1 - 2%) Journal of military pharmaco-medicine n02-2019 For patients undergoing laparoscopic surgery, CO2 is pumped into the abdomen and maintains a pressure of 10 - 12 mmHg during the surgery, the tumor-containing specimen is removed from the abdominal wall through a small incision that is approximately cm long For opensurgery patients, incision is made about 20 - 25 cm long in the middle abdominal wall Postoperative treatment is administered antibiotics combination of 3rd generation cephalosporine and MDA for days, intravenous nutrition depending on the patient’s ability to recover digestion report of Ohkawa H et al in 1979 [6] The erythrocyte after being collected in the above step will be washed with PBS (phosphate buffer saline), then broken to collect erythrocyte membranes Determination of MDA content by TBA test (based on the reaction between MDA and thiobarbituric acid - TBA), the absorbance of the product is measured at a wavelength of 532 nm Total protein content determined by Bradford method, erythrocyte MDA concentration was calculated and expressed in ng/mg protein * Process of quantifying erythrocyte MDA: Blood samples were taken from the peripheral vein and inserted into the tube containing ethylene diamine tetraacetic acid (EDTA), the blood after taking was stored at 4oC then centrifuged 4,000 rpm to separate erythrocyte and plasma Quantification of erythrocyte MDA based on the procedure was modified from the * Statistical analysis: The results for erythrocyte MDA are presented as mean value ± SD, for other characteristics expressed in proportional form Statistical analysis was performed by Mann-Whitney, Friedman or Wilcoxon tests, with a p < 0.05 value considered statistically significant Data are processed and analyzed by SPSS 20.0 software RESULTS Characteristics of the research group Table 1: Characteristics of clinical pathology and surgical treatment Characteristics n % < 60 25 41.7 ≥ 60 35 58.3 Male 36 60.0 Female 24 40.0 I 11.7 II 27 45.0 III 26 43.3 Age Gender Disease stage 215 Journal of military pharmaco-medicine n02-2019 Laparoscopy 34 56.7 Open 26 43.3 Not expanded 49 81.7 Expanded 11 18.3 60 100 Surgical endoscopic method Surgical level Total The average age of the research group was 60.4 ± 11.3 (the lowest was 29 years, the highest was 87 years), 25 patients were < 60 years of age; 35 patients were ≥ 60 years old Males: 36 patients and females: 24 patients There were patients in stage I (11.7%), 27 patients in stage II (45%), 26 patients in stage III (43.3%) 34 patients underwent laparoscopic surgery and 26 patients underwent open surgery In 11 cases of surgery expansion, there were patients with gastrectomy, patients with small intestine resection, patient with hysterectomy, patient with oophorectomy and the rest had to with total colectomy Changes of erythrocyte MDA index before and after surgery Results of changes in erythrocyte MDA index before and after surgery are presented in table 2, table and figure Table 2: Erythrocyte MDA content at postoperative time MDA content n max Average Median Before surgery 60 0.045 0.624 0.168 ± 0.10 0.145 day after surgery 60 0.070 0.789 0.217 ± 0.13 0.186 days after surgery 60 0.055 0.557 0.190 ± 0.11 0.147 days after surgery 60 0.034 0.726 0.179 ± 0.12 0.152 p* 0.001 (* Friedman test) Table 3: p value when comparing MDA content by time points p* (n = 60) MDA before surgery MDA after day MDA after day 0.001 MDA after days 0.795 0.020 MDA after days 0.985 0.003 (* Wilcoxon test) 216 MDA after days 0.942 Journal of military pharmaco-medicine n02-2019 0,8 protein (ng/mg content MDA Hàm lư?ng MDA (ng /mg protein) 0,7 0,6 0,5 0,4 0,3 0,2 0,1 0,0 day 11 ngày Trư?c m? Before ngày 33 days 77 ngày days Figure 1: MDA variation of study group according to the time of surgery The erythrocyte MDA after day had the highest value, before surgery it had the smallest value, the difference between the times was statistically significant with p < 0.05 Comparing paired difference the erythrocyte MDA values by each time showed the between MDA value before surgery with day, between day after surgery with after surgery days and days, the difference was statistically significant with p < 0.05 The difference of erythrocyte MDA between times: before surgery, days and days after surgery had no statistical significance with p > 0.05 The change in erythrocyte MDA after surgery according to surgical characteristics Table 4: Erythrocyte MDA values at different times by surgery Surgery method Erythrocyte MDA content n p* Laparoscopic (n = 34) Open (n = 26) Before surgery 60 0.174 ± 0.11 0.161 ± 0.09 0.536 day after surgery 60 0.233 ± 0.13 0.197 ± 0.11 0.187 days after surgery 60 0.200 ± 0.12 0.177 ± 0.11 0.408 days after surgery 60 0.200 ± 0.14 0.151 ± 0.09 0.072 0.003 0.043 p** (* Mann-Whitney test; ** Friedman test) 217 Journal of military pharmaco-medicine n02-2019 The erythrocyte MDA value in both laparoscopic and open groups according to the time after surgery, the difference was statistically significant with p < 0.05 However, from time to time, after surgery days, erythrocyte MDA in open surgical group tends to be lower than in laparoscopic group Table 5: Erythrocyte MDA content at the time of surgery Surgical level Erythrocyte MDA content n p* Not expanded (n = 49) Expanded (n = 11) Before surgery 60 0.177 ± 0.11 0.130 ± 0.04 0.221 day after surgery 60 0.229 ± 0.13 0.164 ± 0.08 0.122 days after surgery 60 0.201 ± 0.12 0.141 ± 0.07 0.067 days after surgery 60 0.179 ± 0.12 0.140 ± 0.06 0.24 < 0.001 0.664 p** (* Mann-Whitney test; ** Friedman test) Comparing the postoperative period, in the not expanded surgery group, the MDA change had statistical significance with p < 0.05; in the expanded surgery group, the MDA change did not have statistical significance with p > 0.05 Thus, after the surgery, in the expanded surgical group, the MDA value change occurred less than compared to the not expanded surgery DISCUSSION In colon cancer, the higher oxidative stress status in cancer tissue has been confirmed by many researches [7] So surgery for cancer treatment is a complicated process because although surgery can remove a large part of the oxidative mass of the tumor, surgical injuries can stimulate a stressful response, leading to a release of a large amount of free radicals, disrupting the internal balance of oxidation - antioxidant systems So after radical surgery of colon cancer, oxidative stress changes may increase or decrease Our research results evaluated the oxidative stress through quantitative erythrocyte MDA in peripheral blood showed that after day’s surgery, MDA value increased significantly compared to 218 preoperation (p < 0.05, table 2, table and chart 1) Our results as well as many other researches showed a higher level of immediate oxidative stress after surgery, because any form of injury, including surgery, leads to oxidative stress In abdominal surgery, increased formation of free radicals was also the result of ischemia reperfusion of abdominal organs and inflammation of the body after surgery [8] The results when quantifying erythrocyte MDA at days and days after surgery showed that the level of oxidative stress compared with day after surgery showed signs of reducing (p < 0.05), but compared with preoperation, the difference was not statistically significant (p > 0.05) This showed that after surgery, the body activates the anti-oxidation mechanism and over time, the redox-reduction Journal of military pharmaco-medicine n02-2019 equilibrium tended to be reestablished, but for some patients, days after surgery may not be enough for the body recovery to fight against oxidative stress caused by surgery In laparoscopic surgery, the injection of CO2 gas to maintain abdominal pressure during surgery can increase the formation of free oxygen radicals by inhibiting the activity of SOD, an antioxidant enzyme and increase the level of MDA, a product of lipid peroxidation process [9], but laparoscopic surgery is also known to cause less surgery injury and inflammatory reactions than open surgery, so oxidative stress was lower Some research results on colon surgery also showed that some signs of oxidative stress were significantly reduced in the endoscopic group, compared to the open surgical group [4, 10] Similarly, with the group of patients with expanded surgery, there often was a higher level of surgery, more removed organs, more surgical seizures, and the increase in oxidative stress is also larger However, in our study when comparing at the postoperative time points of changes in erythrocyte MDA between groups of laparoscopic surgery and open surgery; between groups of expanded and not-expanded surgery, the results were incompatible, in the group of laparoscopic surgery and not-expanded surgery, postoperative oxidative stress tended to more highly increase surgery, and on the 7th day after surgery, the erythrocyte MDA tended to decrease REFERENCES K Hospital Report on the National Project on Cancer Prevention in Vietnam in the period of 2008 - 2010 Hanoi Mena S, Ortega A, Estrela J.M Oxidative stress in environmental-induced carcinogenesis Mutation Research 2009, Mar 31, 674 (1-2), pp.36-44 Skrzydlewska E, Sulkowski S, Koda M.B et al Lipid peroxidation and antioxidant status in colorectal cancer World J Gastroenterol 2005, 11 (3), pp.403-406 Arsalani-Zadeh R, Ullah S, Khan S, MacFie J Oxidative stress in laparoscopic versus open abdominal surgery: A systematic review J Surg Res 2011, 169, pp.59-68 Lauschke H, Tolba R, Burger B et al Lipid peroxidation as additional marker in patients with colorectal cancer European Surgical Research 2002, 34 (5), pp.346-350 Ohkawa H, Ohishi N, Yagi K Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction Analytical Biochemistry 1979, 95, pp.351-358 Chang D, Wang F, Zhao Y.S, Pan H.Z Evaluation of oxidative stress in colorectal cancer patients Biomed Environ Sci 2008, 21, pp.286-289 Thomas S, Balasubramanian K.A Role of intestine in postsurgical complications: Involvement of free radicals Free Radic Biol Med 2004, 36, p.745 CONCLUSION Sare M, Yilmaz I, Hamamci D et al The effect of carbon dioxide neumoperitoneum on free radicals Surg Endosc 2000, 14, pp.649-652 In patients with colon cancer who underwent radical surgery, compared to preoperation, the erythrocyte MDA was significantly increased at day after 10 Tsuchiya M, Sato E.F, Inoue M et al Open abdominal surgery increases intraoperative oxidative stress: Can it be prevented? Anesth Analg 2008, 107 (6), pp.1946-1952 219 ... highly increase surgery, and on the 7th day after surgery, the erythrocyte MDA tended to decrease REFERENCES K Hospital Report on the National Project on Cancer Prevention in Vietnam in the period... peroxidation, is widely and popularly applied in many studies Therefore, the aim of this research is: To evaluate the change of erythrocyte MDA in peripheral vein in patients with colon cancer after radical. .. pharmaco-medicine n02-2019 Radical surgery is still the main method for treating colon cancer, but the trauma in surgery also causes oxidative stress for the body due to the release of many free radicals The