Objective To establish a human breast cancer cell line with green fluorescent protein(pAcGFP)gene.Methods Human breast cancer cell lines MCF-7 were transfected with pAcGFP-C1 by Fugene HD Transfection Reagent.The cells which express the pAcGFP gene were selected by geneticin(G418).The expression of pAcGFP were detected by inverted fluorescence microscope and flow cytometry,the growth curve and invasive power were observed to compare the biological behavior of the cells transfected with pAcGFP and untransfected ones.Results A human breast cancer cell line with pAcGFP was established,named MCF7-pAcGFP.The transfection rate was 85%.The growth curve and invasive power of the transfected cells were similar to untransfected ones(P＞0.05).Conclusion The MCF7-pAcGFP cells could stably express AcGFP,and pAcGFP transfection does not change the biological behavior of the cells.

Objective To investigate the effects of arsenic trioxide on angiogenesis of hepatic VX2 tumors in rabbit.Methods 16 hepatoma models were reproduced with VX2 cell lines in rabbits and then randomly divided into trial group and control group,each group had 8 rabbits.Sodium chloride was injectd in control group and arsenic trioxide was injected in trail group by the peripheral vein of rabbit ears for 3days respectively.The intratumoral microvessel density(MVD)and the expression of vascular endothelial growth factor(VEGF)were evaluated by immunohistochemical staining.Results The expression of MVD(P=0.021＜0.05)and VEGF(P=0.012＜0.05)in trail group were significantly lower than those of control.A significant positive correlation between MVD and VEGF was found in both trial and control group.Conclusion Arsenic trioxide can inhibit the angiogenesis of hepatic VX2 tumors in rabbit obviously.

Objective BRCA1 has been known as breast cancer susceptibility genes which is closely associated with familial breast cancer and early onset breast cancer.The objective of our research is to investigate the prevalence of BRCA1 gene mutations among HeiLongJiang patients with familial or at early onset breast cancer.Methods 92 breast cancer patients with familial history or have suffered from breast cancer at the age of less than 35 years old were analyzed.A comprehensive mutation analysis(except exon1 and exon4)was performed through SSCP and subsequent DNA direct sequencing.Results In 92 breast cancers,five mutations were identified that included 3 novel mutations and 2 previously reported mutations which are all nonsense mutation.Conclusion The BRCA1 may play a very important role in familial breast cancer of HeiLongJiang population.Three novel mutations in BRCA1 may be mutations characterized to familial breast cancer of Chinese HeiLongJiang population.The mutation frequency of BRCA1 in familial breast cancer patients from HeiLongJiang is obviously lower than that in the western countries.However,the nonsense mutation 3712insG were detected in three patients which indicated that this mutation may be a founder mutation.

Objective To evaluate the clinical diagnosing value in the differentiation of prostatic carcinoma by three-dimensional power Doppler.Methods The characteristics of blood flow examined by three-dimensional power Doppler were analyzed retrospectively and compared pathologically in 68 prostatic lesion.Results Vascularity index was significant differences between prostatic benign and malignant lesion(t= 4.654,P=0.000).Using VI,the sensitivity and specificity were 77.8%and 65.2% respectively in diagnosing prostatic carcinoma.VI in well,moderate,poorly differentiated prostatic carcinoma was 0.0975±0.0680,0.1811±0.0977 and 0.2588±0.0577,respectively,and there were significant differences among groups(P＜0.000).Conclusions Three-dimensional power Doppler offers a new way in diagnosis of prostatic carcinoma,and is helpful to the differentiation of prostatic carcinoma.

Objective To determine acute toxicity reactions and approximate lethal dose of the anti-tumor drug of Chelerythrine.Methods The drug toxicity and side effects were observed when the mice,rat and canine were treated according to toxicological procedures of safety evaluation for drugs.Results The animals were quiet and unanimated,breath was deep and slow,its fur was thin and lackluster;the rats were not sensitive to feeble stimulations.The dead animals were anatomized and abnormal changes were observed:liquid with blood cells in thorax,tumescence of many organs with congest and ecchymosis.A delayed bleeding time was observed in the pinhole of tail vein.Symptoms of celiac stimulation were also observed in rats,included the wring reaction and breadthwise welter.The median lethal dose(LD₅₀)was 22.84mg/kg for mouse iv and 24.30mg/kg for rats ip.Approximate lethal dose was 19.94mg/kg for Beagle canine iv.Conclusions The drug on animals have significant toxicity,the cause of death related with the liquid into the blood caused by vascular dilation,congestion,multiple organ blood circulation disorders,acute respiratory failure caused by respiratory disorders and direct stimulation of abdominal organs.

Objective To observe the effects of Ursolic acid(UA)on adhesion,invasion and migration of HO-8910PM cell line in vitro,and discuss its possible mechanism.Methods Effect of UA on adhesion of HO-8910PM cells were determined with MTT assay,the effects of UA on invasion and migration of HO-8910PM cells were evaluated using Transwell chambers attached with polycarbonate filters and reconstituted basement membrane.The exprssion of MMP-2 mRNA,MMP-9 mRNA was measured by RT-PCR in HO-8910PM cells,the protein expression of MMP-2,MMP-9 was analyzed by Western blot.Results In the cell-Matrigel adhesion experiment,inhibition rate of UA groups at different concentrations of 10,20,30μmol/L for 24h were 44.03%,46.62% and 55.11% respectively which were significantly higher than those of control groups(P＜0.05),after 24 hours,the ability of the different concentrations of UA treated HO-8910PM cells to invade the reconstituted basement membrane decreased significantly(P＜0.01).UA can also inhibit HO-8910PM cells migration(P＜0.05).RT-PCR and Western blot results showed decreased expression of MMP-2,MMP-9 in HO-8910PM cells treated with(10,20,30μmol/L)UA for 24 hours.Conclusion The inhibition of invasion ability of HO-8910PM cells by UA may be attributed to decreases of the expression of MMP-2,MMP-9.

Objective To explore the correlation of chicken skin mucosa beside adenoma and adenoma canceration tissue,debate clinical significance of chicken skin mucosa beside colorectal carcinoma.Methods To observe the expression of Survivin,COX-2 and Ki-67 in 90 cases(chicken skin mucosa beside adenoma 35 cases;adenoma canceration tissue,30 cases;normal colorectal mucosa 25 cases)by immunohistochemistry staining.Results The expression of Survivin was 0 in normal colorectal mucosa,and the expression of Survivin in chicken skin mucosa by the side of adenoma and adenoma canceration tissue was non statistically significant(P＞0.05);The expression of Ki-67 and COX-2 in normal colorectal mucosa,chicken skin mucosa by the side of adenoma and adenoma canceration tissue was positive,non statistically significant between fore two groups(P＜0.05),statistically significant between last two groups(P＞0.05)statistically significant between last two groups(P＞0.05).Conclusion Chicken skin mucosa by the side of colorectal adenoma maybe primarily lesion precancerous and a risk factor of colorectal adenoma canceration,should be early treated by the operation of electric coagulation and electroblation colorectal adenoma.

Objective To study the mechanism of TP inducing apoptosis of colon cancer SW-480 cell.Methods The influce of TP on proliferation of SW-480 cell in vitro was detected with MTT method;The ultrastructure of cell was observed with electron microscopy.Results Compared with control group,the proliferation of SW-480 cell lines in experimental group was obviously inhibited,spindle-shaped and round cells loosely distributed,some round cells increased within the particles after TP treatment.Cell sizes,nuclear atypia,and a collapse of apoptosis,nuclear biased towards one side,the cytoplasm is rich in free ribosome,mitochondria thick,the cell apoptotic body were seen under electron microscope.Conclusion TP can inhibit the growth of SW-480 cell and induce cell apoptosis in vitro.

Objective To investigate the clinicopathologic features of gastric cancer with peritoneal dissemination,and also to evaluate the impact of different operations on the prognosis.Methods A statistical analysis was performed retrospectively with regard to clinicopathologic features and survival of 91 cases of gastric cancer with peritoneal dissemination.Results Univariate analysis revealed that tumor size,tumor location,degree of differentiation,Borrmann types,organ infiltration,lymph node metastasis,depth of invasion and hepatic metastasis were correlated with Peritoneal dissemination(P＜0.05).The Logistic regression model revealed that the depth of invasion and organ infiltration were independent factors(P＜0.05).The difference of age,ascites and organ infiltration between the non-radical resection and palliative operation groups was statistically significant(P＜0.05).The different survival time of palliative operation group with feeding neostomy,gastrointestinal anastomosis and exploratory laparotomy was not statistically significant(P＞0.05).The median survival time in non-radical resection group was 13.4 months and in palliative operation group was 4.2 months.The difference was statistically significant(P＜0.01).Multivariate analysis by Cox regression indicated that the type of operation and tumor size were significant factors affecting the prognosis of the patients with peritoneal dissemination.Conclusions The clinicopathologic features of patients with gastric cancer are intimately correlated with peritoneal dissemination.Non-radical resection can improve the prognosis of peritoneal dissemination patients of gastric cancer.

Objective To investigate the effects of transcutaneous electrical acupoint stimulation(TEAS)on perioperative serum levels of cytokines and postoperative analgesia in patients undergoing radical mastectomy.Methods Forty(ASAⅠ/Ⅱ)female patients undergoing radical mastectomy were randomly divided into two groups(n=20 each).In group E,patients received epidural blockade only.In group H,patients received TEAS plus epidural blockade.In group H,electrical stimulations of bilateral Hegu acupoints(LI4)and Neiguan acupoints(P6)were started from 30 minutes before epidural blockade and continued until the end of operation.Blood sample were taken before(T₀)and the end of(T₁)and 12h(T₂)after operation for determination of serum IL-6,IL-10 and cortisol concentrations in both groups.The postoperative VAS were recorded.Results The two groups were comparable with respect to age,body weight,duration of operation and the amount of fluid infusion and blood loss.Serum cortisol concentration was significantly lower at T₁ and T₂ in group H than that in group E(P＜0.05).The serum IL-6 concentration was significantly increased after operation as compared with baseline in both groups and was significantly lower in group H than that in group E(P＜0.05).The serum IL-10 concentration was significantly higher at T₂ in group H than that in group E(P＜0.05).During 12 hours after operation,VAS of group H was lower than that of group E(P＜0.05).Conclusion TEAS may attenuate cortisol and IL-6/IL-10 responses after surgery.TEAS also has analgesic effect in patients undergoing radical mastectomy.

Objective To explore the value of 3-dimensional digital subtraction in the diagnosis and treatment of the obstructive jaundice caused by the malignant tumours.Methods Twenty patients with obstructive jaundice caused by the advanced stage of malignant tumours were examined with 3-dimensional digital subtraction after the the Ultrasonic Guide Puncture intrahepatic bile duct extension,injecting the contrast material.Then the 3-dimensional imaging on the inspection results were conducted and pathological changes were observed.At the same time,draw another twenty medical case of this kind of terminal disease without 3-dimensional digital subtraction images in the corresponding hospitalized treatment period randomly.Next,conduct the comparison and analysis of the observation data with the two groups of patients.Results 3-dimensional digital subtraction images demonstrated the position,precision length of the bile duct malignant stricture,the shape of the stricture part of the pathological changes clearly,which contributes to the precise localization and the selection of suitable treatment method.And through the comparison,we found that the contrast material and total radiation dosage had been reduced and the operation process had been shortened distinctly in the cases with 3-dimensional digital subtraction images.Conclusion The 3-dimensional digital subtraction can reduce the operation time,decrease the dosage of contrast material and radiation.It plays an irreplaceable role in the diagnosis of the obstructive jaundice caused by the malignant tumours,in the precise measurement of the pathological changes,in the selection of the suitable therapy.

Objective This study is to quantitatively analyse the set-up error in the different positions of the irradiated patients using electronic portal imaging device(EPID)to provide the set-up accuracy in the different positions and to understand the acceptance between different oncologist.Methods From January 2006 to December 2007,58 patients were involved in this study,who randomly selected from the different oncology treatment group by different positions(20 patients in the head and neck,19 patients in the chest,19 patients in the abdomen and pelvic).The treatment of all cases were implementated by Elekta Precise accelerator,scanned by Philips AcQ-Sim CT and/or simulated by Nucletron Simulix-HQ simulator,planned by ADCC Pinnacle3 7.4 system.Treatment planning computed tomography(CT)scans were used to create orthogonally digitally reconstructed radiographs(DRRs)for portal image registration,which were compared with portal images from an electronic portal-imaging device(EPID)of Elekta′s iView GT by comparing bone structure in horizontal and vertical directions.Results A total of 102 data were reviewed,the set-up error of horizontal position is 1.02±1.15mm(range 0-5mm),the vertical position is 1.11±1.20mm(range 0-6mm).More than 90% setup error is less than 3mm.The smallest setup error in the horizontal direction and vertical direction is 0.64±0.65mm and 0.91±0.98mm respectively with head or head and neck fixed.The set-up errors were different between the different oncologist.Conclusions The majority of setup errors is in the permissible range.Need to improve the recognition of the EPID in the chest,abdominal pelvic anatomy.The implementation of the X-ray simulator before treatment is an effective means to reduce setup error.

Objective To investigate the relation between the expression of matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 14(MMP-14)in breast cancer and tumor size,differentiation and lymph node metastases.Methods Sixtyeight cases of breast cancer and twentyfour cases of breast benign disease were studied by PV Immunohistochemical staining with anti MMP-2,MMP-14 monoclonal antibodies.Results 1.The expression rates of MMP-2 and MMP-14 were 55.9 %(38/ 68)and 58.8 %(40/68)in breast cancers respectively,they were 25.0%(6/24) and 16.7%(4/24)respectively in breast benign disease.The expression rates were significantly higher than those in breast benign disease(P＜0.05);2.Expression of MMP-2 and MMP-14 in breast cancers were positive correlated with axillary lymph node metastases and tumor size(P＜0.05).Conclusion The expression of MMP-2 and MMP-14 closely correlated to lymph node metastasis and tumor size,they can act as prognostic markers of invasion and metastasis for patients with breast cancer.