| BackgroundPrimary liver cancer,especially hepatocellular carcinoma(HCC),its incidence is higher,global 1 million new cases each year,early clinical surgical resection can to a certain extent,prolong the survival of patients with primary liver cancer[MP1],but in clinical,only 10%to 30%of patients with liver cancer surgery can be done,the rest of the majority of patients,due to the location of the tumor,the degree of development of transfer reason cannot undergo surgery and pathology classification,this part of patient treatment effect is poor,mostly in advanced disease lead to liver failure.Radiofrequency ablation guided by ultrasound technology because of its small trauma,wide adaptation,has the certain curative effect,wide to treat cancer,in has certain clinical application and clinical studies have found that radiofrequency ablation guided by ultrasound technology of liver cancer have certain control effect,to a certain extent,ease the symptoms of patients.Guided by ultrasound with radiofrequency ablation of liver cancer can cause cancer cells to injury(its main principle is through the high frequency wave cause cancer cells to shocks generate heat,so as to kill cancer cells).Radiofrequency ablation for hepatocellular carcinoma has good curative effect,immune response is small,not easy to relapse,patients with high sensitivity,etc.Clinical radiofrequency ablation guided by ultrasound imaging in more melting the unipolar or multipolar electrode targeted to the tumor site,melting operation under computer control the pulse energy pathway to the tumor tissue,tumor tissue coagulation necrosis under high temperature.High-frequency electromagnetic waves that tumor tissue frequency oscillation and generate heat,heat together temperature than the extent of the cancer resistance,high temperature damage leading to cell death,usually arrived at 70-100 ?,the temperature can lead to irreversible cell necrosis.But how radiofrequency ablation associated with tumor cell necrosis,how to suppress tumor development and metastasis,its specific mechanism is unclear.And the growth of tumor in transfer of the main and essence in capillary tumor growth,more[MP2]scholars believe that the faster the essence in capillary tumor growth,its malignant tumor and the rate of transfer is more bigger.Therefore,at present most scholars use the tumor microvascular density index to reflect the possibility of tumor malignant degree and metastasis,inhibit the growth of tumor blood capillary can be a new treatment.So this article will explore ultrasound ablation by animal experimental rabbit liver cancer after residual cancer molecular biology behavior,for the clinical treatment of local tumor recurrence after ultrasonic ablation foundation,in order to prolong the lifetime.HIF to hypoxia inducing factor(hypoxia inducible factor,HIF),including its family HIF-1 alpha,beta,HIF HIF-1-2 alpha,HIF-2 beta,HIF-3 alpha,hypoxia induced factor is in the early 90 s found that americans have trace expression in normal environment,trace the expression of hypoxia inducing factor in/MP4 oxygen dependence under the action of ubiquitin proteasome degradation immediately,anoxic condition of oxygen induced factor of agglomeration,more stable.Thus hypoxia inducing factor detection can indirect reaction cell or organism hypoxia state.Vascular endothelial growth factor(vascular endothelial growth factor,VEGF)is very important in the process of tissue vascular growth factors,can promote the growth of endothelial cells division,promote the migration of endothelial cells forming,increasing vascular permeability and endothelial cell activity,so the detection of VEGF can reflect tissue blood vessel growth state,is also the target of tumor therapy.Metal matrix proteins-9(matrix metalloprotein,MMP-9[MP6 plays])for the members of the family of metal matrix protein,its main function is through enzyme secreted by the cell in the form of the original to the outside of the cell,including MMP-9 can through the release of VEGF involved in the vascular endothelial cells,therefore detection of MMP-9 to reflect angiogenesis condition has the vital significanceThe first part:The tissue block preparations method to build a tumor-burdened rabbit liver cancerObjectiveThrough the tissue block preparetions method made a tumor-burdened rabbit liver cancer,lay the ideal animal model for the further experiments.MethodsThis research mainly adopts randomized selection method provided by the Inner Mongolia medical university animal center of clean level 110 New Zealand white rabbit,6 to 7 months of age,body mass of 2300 grams,unlimited male and female.Will be provided by the Beijing union medical college of VX2 tumor cells in 5 rabbit thigh lateral subcutaneous,and extend,14 to 28 days after the thigh growing tumors in tumor growth,tumor tissue and keep incision,step by step a sterile environment will be cut into 1 mm3 tumors had tissue block,and placed in 0.9%saline,tumor tissue suspension packing after grinding,first-80 degrees freezer for 24 hours,and then placed in liquid nitrogen preservation,105 random will be regular fixed rabbit ear margin after intravenous anesthesia in operation stage,the right side of the rib arch routine operation,step by step a exposed abdominal cavity,show the liver,with the eye surgery forceps inclined to wear into the liver,and place the above operation represented tumor grain into the puncture in the liver,with gelatin sponge tamponade,operation and shut abdominal,step by step a breeding in 60 days on ultrasound tumor growth.In accordance with the requirements for the experimental surgery death,infection,rabbit.Observation experiment rabbit general conditions,with high-end real-time color doppler ultrasonic diagnosis system Logiq-the S8-Face,frequency of the probe is 3.0 to 5.0 MHz,measuring plant tumor diameter size,take five rabbit executed do pathologic biopsy of liver cancer specimens to observe its pathological changes.Statistical analysis,using SPSS 13.0 software to calculate,tumor size with a mean diameter of +/-standard deviation.Results1.105 experimental rabbit,only four infections as a result of the surgery,or in the process of building operation error,or due to unknown causes postoperative death,planting tumor survival after 101,with experimental rabbit liver ultrasound on survival checks when found,including three in the liver area can not only observe hyperechoic regions of the tumor to analyze its errors may be due to the process of operation,or plant tumor tissue pieces of deactivation.2.The remaining 98 live a tumor-burdened rabbit liver cancer,the measurement of tumor size was 2.3 +/-0.32 cm,liver ultrasound examination showed that coated can complete,there is no complete individual case,the tumor appeared substance with high echo,dark ring around a wide,also have individual narrower,it is mainly due to the tumor tissue pushed around small blood vessels to form around blood vessels around),liver nodules posterior acoustic often no obvious change,also have rear local enhancement,individual animal models can also be observed in nearby tissue mass.3.To grow successfully in liver cancer 98 randomly selected 5 only in the rabbit,and processing,the pathological and histological section morphology observed no differences of liver cancer were changes to sample for hepatocellular carcinoma(HCC).Obvious liver pathology organization form,often accompanied by huge nuclear and multi-core tumor cells,the nuclei have different size,shape and dyeing,chromatin is more coarse granular,uneven distribution,nuclear membrane hypertrophy,nucleolus split,nuclear fission is asymmetry,wrapped slurry formed a basophilic.Conclusion1.The success of the research through the tissue mass preparetions method established 98 only build a tumor-burdened rabbit liver cancer.2.Building a tumor-burdened rabbit liver cancer of liver cancer histopathological observation conform to the pathological characteristics of cancer of the liver.3.Building a tumor-burdened rabbit liver ultrasound conforms to liver cancer of liver cancer ultrasound imaging characteristics.4.Successful building model for further research on liver cancer recurrence of radiofrequency ablation curative effect and laid a solid foundation.The second part:The rabbit model of liver cancer after radiofrequency ablationand surgical removal of the residual carcinoma angiogenesis of ultrasoundimaging and blood flow signal correlation studiesObjctiveThis research explored mainly through the animal experiment study continuous dynamic observation of rabbit liver cancer after ultrasonic ablation capillaries of residual carcinoma(ultrasound imaging observation)from morphology and blood flow signal measurement,from ablation of liver cancer recurrence after morphological structure on the research process,for the local tumor recurrence after ultrasonic ablation of laying a foundation for clinical treatment.MethodsFirst part of the experiment build the successful tumor-burdened rabbit liver cancer randomly selected from 90,divided into three groups,9 groups,the radiofrequency ablation group of 10,radiofrequency ablation group of 40,then choose different time after cure to radiofrequency ablation group,0 hours after treatment group(10);3 days after treatment group(10);7 days after the treatment group(10)and 14 days after treatment group(10).Surgical removal of the residual tumor group and 40 10%,according to the different treatment time to be grouped,0 hours after surgery group(10);Three days after surgery group(10);7 days after the surgery group(10)and 14 days after the surgery group(10).Insert electrodes radiofrequency ablation group liver area,by a 5 w to 50 w,15 minutes,impedance reached 100%,automatic shutdown,pull out the needle,and stitch the abdomen,and give the anti-inflammatory treatment.Caused the remaining 10%tumor surgery group for regular liver tumor resection of the remaining 10%,suture the abdominal wall,anti-inflammatory therapy.Three groups of rabbits in 0 days after treatment,3 days,7 days,14 days by contrast-enhanced ultrasound to observe the residual carcinoma of the dynamic changes of the microvascular generation,be assured of contrast-enhanced ultrasound in the process of each rabbit check all parameters unchanged(mechanical index,gain and depth),tumor size,ultrasound blood flow velocity and classification,according to figures from low to high divided into 0,1,2,3 level 4,level 0,blood flow signals detected in tumor tissues,blood flow activity level is zero;Level 1,tumor tissue blood flow to the star point short signal,the distribution is less,appear from time to time,and sometimes disappear;Level 2 continuous lines exist in the tumor tissue blood flow signals,has certain continuous flow activities;Level 3 with bulky blood vessels in tumor tissues,blood flow quickly.Contrast agents in U.S.Albunex contrast agents Albunex production company,its dosage reference manual.SUTTER rf radio frequency instrument for Germany therapeutic apparatus,the needle electrode model for bipolar RFA therapy system,the largest rf power for 470 KHZ,host power output of 50-250 wResultsBefore the radiofrequency ablation treatment and surgical excision,grade of microvascular blood flow velocity,the control group 10 cases,vascular level 0 2 cases,blood vessel of grade 1 level 3 cases,vascular level 2 level in 2 cases,blood vessel level for level 3 for 3 cases;Radiofrequency ablation 40 cases,vascular level 0,8 cases of blood vessels of grade 1 level 14 cases,vascular level 2 to 5 cases,the blood vessels of grade 3 to 13 cases;Surgical removal of the remaining 10%group of 40 cases,vascular level 0 9 cases,13 cases of vascular level for level 1,level 2 vessels for 6 cases,the blood vessels of grade 3 to 12 cases,by the analysis of variance,P>0.05,no statistically significant difference between the three groups.After radiofrequency ablation for pathological observation,operation area most tissue necrosis,nuclear pyknosis,agglutination,dark blue,chromatin cleft,debris,nuclear membrane dissolved,not the nucleus outline,swelling of liquefaction in interstitial cells.After 7 days and 14 days after radiofrequency ablation area in liquefied,necrotic range further expand,finally have to observe a fibrosis around the liquefaction area.Ablation for ultrasound imaging examination,after check immediately after ablation in ablation area is seen here surrounded and infiltrated by gasification echo,boundary shape is fuzzy,the resolution is not clear,the blood flow signals become a level 1,basic undetectable blood flow,ablation of postoperative 3 days before the tumor size and ablation boundary,fuzzy boundaries,the ablation area in high echo,blood flow signals become a level 1,basic undetectable blood flow.7 days to 14 days for ultrasound examination,boundary gradually clear,high ablation area appeared echo or high mixed echo,blood flow signals become 2-3.Grade of microvascular blood flow velocity,the control group in 10 cases,vascular level 0 2 cases,blood vessel of grade 1 level 3 cases,vascular level 2 level in 2 cases,blood vessel level for level 3 for 3 cases;Radiofrequency ablation group of 10 cases,0 days vascular level 0 7 cases,the blood vessels of grade 1 level in 2 cases,blood vessel level 2 for 1 case,vascular level to level 3 to 0 cases;Radiofrequency ablation group of 10 cases,3 days blood vessel level 0 5 cases,the blood vessels of grade 1 level 3 cases,vascular level 2 level in 2 cases,blood vessel of grade 3 to 0;Radiofrequency ablation group of 10 cases,7 days of vascular level 0 4 cases,vascular level 1(3 cases),vascular level 2 level in 2 cases,blood vessel level for level 3 in 1 case;14 days radiofrequency ablation group of 10 cases,vascular level 0(3 cases),vascular level 1 in 4 cases,vascular level 2 level in 2 cases,vascular level for level 3 in 1 case.For surgical removal of the residual tumor group 10%,0 days group of 10 cases,vascular level 0,8 cases of blood vessels of grade 1 class 1 case,vascular level 2 for 1 case,vascular level to level 3 to 0 cases;For surgical removal of the residual tumor 10%3 days group of 10 cases,vascular level 0 to 7 cases,the blood vessels of grade 1 level in 2 cases,blood vessel level 2 for 1 case,the blood vessels of grade 3 to 0;For surgical removal of the residual tumor 10%7 days group of 10 cases,vascular level 0 6 cases,the blood vessels of grade 1 level 4 cases,blood vessel level 2 to 0 cases,blood vessels of grade 4 to 0;Radiofrequency ablation group of 10 cases,14 days blood vessel level 0 6 cases,the blood vessels of grade 1 level 4 cases,blood vessel level 2 to 0 cases,blood vessels of grade 4 to O.By analysis of variance,F =2.35,P<0.05,the difference between groups were statistically significant,we will be the same time the radiofrequency ablation group compared with 10%residual cancer surgery group two,P<0.05,differences were statistically significant,radiofrequency ablation group blood levels lower than the control group,but higher than 10%surgical removal of the residual carcinoma group.Contrast-enhanced ultrasound showed that radiofrequency ablation,around the liver cancer development increases rapidly,lasts for 5 to 8 seconds,and then developing around 20 seconds and then disappear;Melting,0 days after 3 days,after injection of contrast agents for 15 seconds,a blood flow signal around the tumor,liver area appear defect area,internal to see blood flow signal;Melt after 7 days,14 days,before defect appeared strengthen blood flow,blood flow to the group of 7 days,2 cases with blood flow to strengthen,14 days group with 4 cases of blood flow to strengthen foci.Melting,0 days after 3 days,after injection of contrast agents for 15 seconds,a blood flow signal around the tumor,liver area appear defect area,internal to see blood flow signal;Melt after 7 days,14 days,before defect appeared strengthen blood flow,blood flow to the group of 7 days,2 cases with blood flow to strengthen,14 days group with 4 cases of arterial strengthening focal;0 10%residual cancer surgery group day,3 days,after injection of contrast agents for 15 seconds,a blood flow signal around the tumor,liver area appear defect area,internal to see blood flow signal;10%residual cancer surgery group after 7 days,14 days,before the defect appears strengthen blood flow,blood flow to the group of 7 days,1 cases of blood flow to strengthen,14 days group with 2 cases of blood flow to strengthen foci.And comparing two group,P<0.05,differences were statistically significant.Conclusion Radiofrequency ablation in part can kill tumor foci,but as a prolonged postoperative ultrasonic blood level is higher than 10%residual cancer surgery group.2.Radiofrequency ablation in part can kill tumor foci,but as a prolonged postoperative ultrasonic angiography strengthen blood flow is higher than 10%residual cancer surgery group.The third part:the rabbit model after radiofrequency ablation and surgical removal of the blood supply of liver cancer classification relationship with angiogenesis researchObjectiveThis research mainly through the animal experiment to investigate the ultrasound ablation after the relationship between MVD and VEGF by continuous dynamic correlation research,the molecular mechanism research on liver cancer recurrence after ablation process,for the local tumor recurrence after ultrasonic ablation of laying a foundation for clinical treatment.MethodsThe second part of the packet radio frequency control,radiofrequency ablation and surgical removal of the remaining 10%rabbit according to the selected point in time,to be put to death the residual carcinoma model rabbit,tumor tissue samples,to be part of the pathological tissue for cancer of the liver of microvascular density(MVD)staining and count,then the other pathological tissue liquid nitrogen preservation and paraffin embedding,respectively.Microvascular count the microvascular density of dyeing method proposed reference to Wendy,interstitial cells dyed buffy considered positive cells,cell mass near positive staining for blood vessels,namely the number of blood vessels at low magnification observation technology,usually choose three concentrations of blood vessels,count after calculating mean +/-standard deviation.Will organize the specimen immunohistochemical staining method mainly refer to the instruction with rabbit polyclonal antibody staining for VEGF,MMP-9 expression.Pathological tissue slice after embedding with PBS wash three times,hydrogen peroxide block 10-15 minutes,using high-temperature high-pressure antigen of repair,repair complete cooling reoccupy PBS solution is rinsed three times,can be closed 15 minutes and 26 degrees Celsius incubation,serum respectively and VEGF,MMP-9 a fight,37 degrees Celsius incubation reoccupy PBS solution is rinsed three times after 2 hours,and low resistance,PBS wash again after incubation at room temperature for 15 minutes,again add SP solution and DAB solution,the former add after place 15 minutes,reoccupy PBS wash three times,the latter DAB chromogenic wood dyeing after water rinse,the microscopic observation color.Using rabbit polyclonal antibody of VEGF,MMP-9 to join the pathological tissue section continuous culture for 1 hour at room temperature,then add in the substrate(Abcam,Cambridge,UK),1-5 minutes with wood grain to after dyeing,dyeing and sealed with cover glass after dehydration.Immune histochemical method result determination,the main reference to VEGF,MMP-9 test kit,including VEGF,MMP-9 are located in the cytoplasm,low power view,positive cells of cytoplasm tan dye particles,dyeing range is more than 10%,considered to be positive,less than 10%,considered to be negative.ResultsImmunohistochemical detection:the radio frequency(rf)group are in high expression of the VEGF,MMP-9(67.3%),respectively(63.5%).Rf group 0 days VEGF,MMP-9 positive expression rate of(15.3%),respectively(23.6%);3 days after radiofrequency group of VEGF,MMP-9 positive expression rate of(16.7%),respectively(26.5%);7 days after radiofrequency group of VEGF,MMP-9 positive expression rate of(34.9%),respectively(37.3%);14 days after radiofrequency group of VEGF,MMP-9 positive expression rate of(57.6%),respectively(61.5%);Surgical removal of the remaining 10%0 days VEGF,MMP-9 positive expression rate of(16.8%),respectively(22.7%).Surgical removal of the remaining 10%after 3 days group of VEGF,MMP-9 positive expression rate of(15.2%),respectively(23.6%);Surgical removal of the remaining 10%7 days group of VEGF,MMP-9 positive expression rate of(26.6%),respectively(28.6%).Surgical removal of the remaining 10%14 days set of VEGF,MMP-9 positive expression rate of(35.2%)(39.6%)respectively.By the analysis of variance,F = 4.56,P<0.05,the difference between groups were statistically significant,we will radiofrequency ablation group under the same time,compared with 10%residual cancer surgery group two(P<0.05,differences were statistically significant,radiofrequency ablation group of immunohistochemical score lower than the control group,but higher than 10%surgical removal of the residual carcinoma group.Control group,the blood supply classification and expression of VEGF,MMP-9 and MVD value positive correlation,correlation coefficients were 0.521,0.768,0.814,P is less than 0.05,radiofrequency ablation and surgical residue 10%group every time period blood supply classification and expression of VEGF,MMP-9 and MVD value are related,namely with increasing blood supply grading,the expression of VEGF,MMP-9 and MVD value rise;The radiofrequency ablation group,the expression of VEGF,MMP-9 over time(7 and 14)the expression of VEGF,MMP-9 and MVD values were greater than 10%residual operation.Conclusion1.The control group,radiofrequency ablation and surgical residue 10%group ultrasonic blood flow classification associated with angiogenesis factor,the expression of VEGF,MMP-9 and the technology of microvascular MVD values were positively correlated.2.Radiofrequency ablation group than 10%residual tumor recurrence occurred more often.3.Clinical for liver cancer recurrence after radiofrequency ablation by ultrasonic image classification combined with blood flow to angiogenic factor comprehensive judgement in order to guide the prognosis.The fourth part:the rabbit model of liver cancer after radiofrequency ablation and surgical resection INF 1 alpha,VEGF,MMP-9 mrna and protein expression and blood supply classification correlation studies ObjectiveThis research mainly through the continuous dynamic observation of the animal experiment to investigate the ultrasound ablation HIF-1 alpha,VEGF,MMP-9 mrna and protein expression and blood supply grading correlation research,the molecular mechanism research on liver cancer recurrence after ablation process,for the local tumor recurrence after ultrasonic ablation of laying a foundation for clinical treatment.MethodsMove each IF liver cancer tissue samples-1 alpha,VEGF,MMP-9 mrna and protein expression detection.The separation of RNA and real-time fluorescent quantitative PCR(RT-qPCR)for every 100 mg white rabbit liver cancer tissue samples first in 1 ml of Trizol reagent uniformity,the total mRNA separation using Trizol reagent(Invitrogen,Carlsbad,CA,USA).Redissolved RNA,simple dry RNA precipitation,partly dissolved RNA samples,in the absence of RNA enzymes of the water beat five times,and in 55 ? to 60 ? incubation for 10 minutes,reoccupy amine to dissolve and preservation in 100%formaldehyde-70 ?.Real-time fluorescent quantitative PCR(RT-qPCR)using real-time fluorescent quantitative PCR to alpha,VEGF,MMP-9 HIF-1 in the expression of mRNA level quantitatively,the experimental use SYBR PrimeScript RT-qPCR kit(TaKaRa,Tokyo,Japan),and using the beta actin as the inside.HIF-1 alpha primers for 5'-GTCGGACAGCCTCACCAAACAGAGC3''(justice)and 5'-GTTAACTTGATCCAAAGCTCTGAG-3 '(antisense VEGF primers for 5'-CCCTGATGAGATCGAGTACATCTT-3 '(justice)5'-ACCGCCTCGGCTrGTCAC-3 '(antonym);MMP-9 primers for 5 '-CGGCCACGAGGAACAAACT-3' '(justice),and 5'-CGGAGCACGGAGACGGGTAT3 '(antonym)beta actin primers for 5'-TTCTACAATGAGCTGCGTGTG-3 '(justice)and 5'-GGGGTGTTGAAGGTCAAA-3 '(antisense).All primers from Beijing biological engineering co.,LTD.(Shanghai,China)synthesis.HIF-1 alpha,the expression of VEGF,MMP-9 is based on beta actin standards,expressed as compared with contrast ratio increased and as as with delta delta Ct method of calculation.The selected tissue samples of HIF-1 alpha,VEGF,MMP-9 protein levels by protein imprinting test are analyzed.In the tumor samples before protein extraction of homogenization treatment.Homogenization of samples with lysis agent(Pierce,Rockford,IL,USA)collection and dissolved,reference manual,add protease inhibitor mixture(Thermo Fisher Scientific,Waltham,MA,USA).Protein samples with 12%polyacrylamide gel separation,and then transferred to nitrocellulose membrane.HIF-1 alpha,VEGF,MMP-9 and beta actin is through continuous with HIF-1 alpha,VEGF,MMP-9 or beta actin polyclonal antibody on rabbit,a kind of peroxidase(pod)combined with secondary antibody(Sigma,st.Louis,MO,USA)and the electrochemical luminescence detection system(Amersham,Uppsala,Sweden)quantitatively.ResultsRt-PCR results comparison:the radio frequency(rf)group of HIF-1 alpha,are in high expression of the VEGF,MMP-9 respectively(1.2 +/-0.1)(24.9 + 6.9)(0.92 + 0.2);Rf group 0 days HIF-1 alpha,VEGF,MMP-9 positive expression rate was(4.5 + 1.0),respectively(9.8 + 1.4)(0.2 + 0.02);3 days after radiofrequency group of HIF-1 alpha,VEGF,MMP-9 positive expression rate was(4.3 + 0.5),respectively(12.6 + 2.1)(0.3 + 0.04);Group 7 days after radiofrequency HIF-1 alpha,VEGF,MMP-9 positive expression rate was(2.6 + 0.8),respectively(18.4 + 2.4)(0.6 + 0.07);Alpha group 14 days after radiofrequency HIF-1,VEGF,MMP-9 positive expression rate was(1.9 + 0.3),respectively(21.4 + 3.2)(0.7 +0.02);Surgical removal of the remaining 10%0 days HIF-1 alpha,VEGF,MMP-9 positive expression rate was(3.4 + 0.8),respectively(8.8 + 2.1)(0.1 + 0.05);Surgical removal of the remaining 1 Surgical removal of the remaining 10%group 14 days HIF-1 alpha,VEGF,MMP-9 positive expression rate was(2.3 + 0.2),respectively(16.8 + 2.7)(0.5 +/-0.4).By analysis of variance,F = 5.47,P<0.05,the difference between groups were statistically significant,we will be the same time the radiofrequency ablation group compared with two 10%residual cancer surgery group,P<0.05,differences were statistically significant.HIF-1 alpha protein imprinting method results:the control group,VEGF,MMP-9 is highly expressed,respectively(0.13 + 0.01)(372.6 + 23.5)(0.98 + 0.02);Rf group 0 hours HIF-1 alpha,VEGF,MMP-9 positive expression rate was(0.37 +0.02),respectively(213.3 + 56.2)(0.23 + 0.01);3 days after radiofrequency group of HIF-1 alpha,VEGF,MMP-9 positive expression rate was(0.35 + 0.06),respectively(256.1 + 65.3)(0.35 + 0.02);Group 7 days after radiofrequency HIF-1 alpha,VEGF,MMP-9 positive expression rate was(0.23 + 0.02),respectively(248.3+ 31.5)(0.52 + 0.01);Alpha group 14 days after radiofrequency HIF-1,VEGF,MMP-9 positive expression rate was(0.18 + 0.03),respectively(268.3 + 45.7)(0.78 +/-0.02).Surgical removal of the remaining 10%0 days HIF-1 alpha,VEGF,MMP-9 positive expression rate was(0.32 + 0.08),respectively(196.5 + 23.6)(0.18 +0.05);Surgical removal of the remaining 10%3 days group of HIF-1 alpha,VEGF,MMP-9 positive expression rate was(0.33 + 0.06),respectively(223.4 + 19.8)(0.24+ 0.03)for 7 days after surgical removal of the remaining 10%of HIF-1 alpha,VEGF,MMP-9 positive expression rate was(0.20 + 0.04),respectively(221.6 +25.8)(0.32 + 0.01);Surgical removal of the remaining 10%group 14 days HIF-1 alpha,VEGF,MMP-9 positive expression rate was(0.21 + 0.02),respectively(235.6+ 47.6)(0.36 +/-0.04).By analysis of variance,F = 4.13,P<0.05,the difference between groups were statistically significant,we will be the same time the radiofrequency ablation group compared with 10%residual cancer surgery group two,P<0.05,differences were statistically significant,radiofrequency ablation group blood levels lower than the control group,but higher than 10%surgical removal of the residual carcinoma group.In the control group,the blood supply with HIF-1 alpha mRNA and protein expression showed a negative correlation,and VEGF,MMP-9 mRNA and protein expression of value are related,the correlation coefficient mRNA were respectively 0.6723,0.521,0.768,0.814;Protein expression are:0.741,0.741,0.852,0.663,and P are less than 0.05,radiofrequency ablation and surgical residue 10%group every time period blood supply and grading and VEGF mRNA and protein expression of MMP-9 were positively correlated,and alpha HIF-1 negative correlation mRNA and protein expression,namely with increasing blood supply grading,the expression of VEGF,MMP-9,the expression of HIF-1 alpha;The radiofrequency ablation group,the expression of VEGF,MMP-9 over time(7 and 14)the expression of VEGF,MMP-9 10%residual values were greater than surgery,difference is statistically significant.Conclusion1.The control group,radiofrequency ablation and surgical residue 10%group ultrasonic blood flow classification associated with angiogenesis factor VEGF,MMP-9 mRNA and protein expression was positively correlated with HIF-1 alpha mRNA and protein expression was negatively correlated.2.Radiofrequency ablation group than 10%residual tumor recurrence occurred more often.3.HIF-1 alpha,VEGF,MMP-9 mRNA and protein expression for guiding clinical liver cancer recurrence after radiofrequency ablation has an important value,also can be used as drug interference targets of recurrent liver cancer. |
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