| Carbon nanoparticles suspension injection(Kanalin)has been widely used in clinical practice,having good lymphatic trace effects on radical correction for malignancies.It has been reported widely in the treatment of thyroid cancer,breast cancer,and gastrointestinal malignancies,but rarely in liver cancer.The research group has been committed to the clinical application of carbon nanoparticles on lymphatic trace effects of liver cancer,and the students have made relevant research on trace effects of carbon nanoparticles on liver cancer resection margins and lymphatic drainage in rats.The test proves that the carbon nanoparticles have significant trace effects on the lymphatic drainage of liver cancer in rats,which can improve the positive rate of liver cancer metastasis lymph node excision in rats;in addition,the comparative research on trace effects of carbon nanoparticles and methylene blue on lymphatic drainage of liver cancer in rabbits also proves that carbon nanoparticles have more obvious effects on black-dyed lymph nodes of the drainage of liver cancer in rabbits than methylene blue,reducing the intraoperative time for lymph node dissection.This experiment is to determine mainly the morphology and structure of chemotherapy-drug-carried carbon nanoparticles carrying chemotherapeutic agents under an electron microscope,in order to compare trace effects of carbon nanoparticles alone and carbon nanoparticles carrying chemotherapeutic agentson lymphatic drainage of liver cancer in rats.Objective: the morphology and structure of carbon nanoparticles alone and carbon nanoparticles carrying various chemotherapeutic agents were shotted by a scanning electron microscope,in order to compare trace effects of carbon nanoparticles suspension alone and carbon nanoparticles suspensioncarrying various chemotherapeutic agents on the lymphatic drainage of liver cancer in rats,so as to provide references for the dissection of metastatic lymph node of liver cancer and chemotherapy targeting lymphoid tissues.Methods:1 Reagent Preparation1.1 Use whirlpool + water bath ultrasound shaking method for preparing5-Fu-carried carbon nanoparticles suspension,and epirubicin-carried carbon nanoparticles suspension;1.2 Carbon nanoparticles suspension,5-Fu injection,5-Fu-carried carbon nanoparticles suspension,and epirubicin-carried carbon nanoparticles suspension were prepared as lyophilized powder with lyophilization;and electron microscopy photographs of carbon nanoparticles alone,5-Fu-carried carbon nanoparticles,and epirubicin-carried carbon nanoparticles were shotted with a scanning electron microscope(FEI Quanta200,USA).2 Animal Modeling and Experimental Grouping Peritoneal injection of Walker-256 cell strains was performed to culture and amplify cancer cells by ascites in rats.Liver cancer models in rats were prepared with the direct injection of highly purified cancer cells obtained by the centrifugal purification of cancerous ascites,and divided into the group A,B and C,with 30 cases in each group.5-Fu-carried carbon nanoparticles suspension,epirubicin-carried carbon nanoparticles suspension and carbon nanoparticles suspension alone were injected into the center of tumor bodies of liver cancer in rats of the group A and B(experimental groups)and group C(control group)by aparotomy,respectively.Black-dyed lymph nodes in the drainage region of liver cancer in the three groups were observed and recorded dynamically with a high-definition camera for 30 minutes after injection.Black-dyed lymph nodes were resected seemingly as radical resection of human liver cancer after stably black-dyed lymph nodes,and the dissection was carried out by 3 groups,including the close range group,moderate range group and distant range group.Lymph nodes near the first and second porta hepatis were set as the close range group,lymph nodes around thehepatoduodenal ligament and hepatogastric ligament were set as the moderate range group,and lymph nodes around the root of the mesentery and abdominal aorta were set as the distant range group,in which,the number and position of resected black-dyed lymph nodes were recorded.The degree of dyeing black in lymph nodes were observed,in which,no changes on the color of lymph nodes were recorded as 0 point;partly black-dyed lymph nodes or lymph nodes gray-dyed were recorded as 0.5 point;and all black-dyed lymph nodes were recorded as 1 point.All resected black-dyed lymph nodes were immersed into the 10% neutral formaldehyde solution for preservation and hematoxylin-eosin(HE)staining after marking;and the number of positive metastatic lymph nodes were recorded through the microscopic observation of lymph nodes on the cancerometastasis.3 Statistical Methods Statistical Product and Service Solutions(SPSS)17.0 was used for statistical analysis.The number of resected black-dyed lymph nodes in the three groups was expressed by mean ± standard deviation(-x ±S).Under the normality and homogeneity of variance,comparison among groups was analyzed with single factor variance analysis,and chi-square test was employed for enumeration data,in which,P<0.05 suggested that differences had statistical significance.Result: Scanning electron microscope photographs were acquired by shotting carbon nanoparticles,5-Fu,epirubicin,5-Fu-carried carbon nanoparticles and epirubicin-carried carbon nanoparticles.A total of 876 black-dyed lymph nodes were resected from the three groups,including 292 resected lymph nodes in the 5-Fu-carried carbon nanoparticles suspension group with the average of 14.83 ± 0.874 ones,275 resected lymph nodes in the epirubicin-carried carbon nanoparticles suspension group with the average of 14.60 ± 1.133 ones,and 309 resected lymph nodes in the carbon nanoparticles suspension group with the average of14.73 ± 0.772 ones.The normality test suggested all P > 0.5,and the data obeyed the normal distribution.Levene's test for homogeneity of variancesindicated P=0.853(P< 0.05),in accordance with the homogeneity of variance.The variance test was used for comparison among the three groups: F=0.045,P=0.956(P>0.05),which demonstrated that no significant differences were noted in the number of black-dyed lymph nodes in the three groups,suggesting that there were no obvious differences in the number of black-dyed lymph nodes in the drainage region of liver cancer in rats with between carbon nanoparticles suspension carrying various chemotherapeutic agents and carbon nanoparticles suspension alone.A total of 292 lymph nodes were resected in the 5-Fu-carried carbon nanoparticles suspension group,involving 107 in the close range group(33.6%),129 in the moderate range group(32.8%)and 56 in the distant range group(33.3%).A total of 275 lymph nodes were resected in the epirubicin-carried carbon nanoparticles suspension group,including 102 in the close range group(32.1%),121 in the moderate range group(30.8%)and 52 in the distant range group(31.5%).A total of 309 lymph nodes were resected in the carbon nanoparticles suspension alone group,involving 109 in the close range group(34.3%),143 in the moderate range group(36.4%)and 57 in the distant range group(34.5%).?2 test suggested P>0.05 in the constituent ratio of positions of black-dyed lymph nodes of the three groups,which demonstrated there were no significant differences in the constituent ratio of the number of black-dyed lymph nodes of the three groups distributed in various positions,suggesting that no obvious differences were noticed in trace effects of lymph nodes in various regions of liver cancer in rats with between carbon nanoparticles suspension carrying various chemotherapeutic agents and carbon nanoparticles suspension alone.The black-dyed lymph nodes were 0.876,0.764 and 0.783 points in the carbon nanoparticles suspension group,epirubicin-carried carbon nanoparticles suspension group and 5-Fu-carried carbon nanoparticles suspension group,respectively;and the results demonstrated that there might be different in the degree of dyeing black in lymph nodes of the three groups,indicating that no obvious differences were revealed in the degree of dyeingblack in lymph nodes of the drainage region of liver cancer in rats with between carbon nanoparticles suspension carrying various chemotherapeutic agents and carbon nanoparticles suspension alone.There were a total of 292 resected lymph nodes in the 5-Fu-carried carbon nanoparticles suspension group,including 7 metastatic lymph nodes with positive HE staining with 2.40% in the positive metastatic rate;there were a total of 275 resected lymph nodes in the epirubicin-carried carbon nanoparticles suspension group,involving 5 metastatic lymph nodes with positive HE staining with 1.82% in the positive metastatic rate;and there were a total of 309 resected lymph nodes in the carbon nanoparticles suspension group,involving 8 metastatic lymph nodes with positive HE staining with2.59% in the positive metastatic rate.Fisher's exact test suggested P>0.05 in the positive metastatic rate of black-dyed lymph nodes of the three groups.The results demonstrated that no significant differences were observed in the detection rate of positive metastatic lymph nodes of black-dyed lymph nodes of the three groups,indicating that there were no obvious differences in the detection rate of positive metastatic lymph nodes with between carbon nanoparticles suspension carrying various chemotherapeutic agents and carbon nanoparticles suspension alone.The above results could suggest that there were no significant differences in trace effects of lymph nodes of liver cancer in rats with between carbon nanoparticles suspension carrying various chemotherapeutic agents and carbon nanoparticles suspension alone.Conclusions: As favorable carriers of chemotherapeutic agents,carbon nanoparticles can adsorb relevant chemotherapeutic agents by its wide ability of superficial adsorption.This experiment has confirmed that carbon nanoparticles suspension carrying chemotherapeutic agents does not influence favorable tropism on lymphoid tissues,as well as trace effects of lymphatic drainage of liver cancer region in rats,which provides new thoughts and methods for more complete dissection of lymph nodes under the direct vision of radical resection of liver cancer and targeted chemotherapy with occult lymphoid tissues in clinical. |
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