Objective To detect the disposition of T cells subset in tumor region and the marginal region of tumor(peri-tumor region)and study the relationship between CD4⁺CD25⁺ regulated T cells(Tr)and local immunity in the process of hepatocarcinoma′s development.Methods CD4⁺CD25⁺Tr cells,CD4⁺T cells and CD8⁺T cells were detected in formalin-fixed paraffin-embedded tissues from 54 cases of HCC tissues and 10 cases of normal hepatocellular tissues by S-P immunohistochemical method.To analyze the level of their expression in these cases and relationships between CD4⁺CD25⁺Tr cells and CD4⁺T cells,CD8⁺T cells,CD4⁺T/CD8⁺T.Results 54 patients with HCC had a higher number of CD4⁺CD25⁺Tr cells in tumor region(5.6208±2.7235)than those of the peri-tumor region(3.8554±1.6018)(P=20.001).The number of CD4⁺CD25⁺Tr cells in tumor tissue were correlated with the size of tumor and seed focus or not,the difference was significant(P＜0.01);The number of CD4⁺CD25⁺Tr cells in tumor tissue was negatively correlated with the number of infiltrated CD4⁺T lympholeukocyte and the ratio of CD4⁺T/CD8⁺T(r=-0.459,P=20.015;r=-0.563,P=20.011),and not with the distribution of invasion CD8⁺T lympholeukocyte(r=-0.48,P=20.072).Conclusions CD4⁺CD25⁺Tr cells is the main immunosupression cell in tumor microenvironment of HCC,they suppress proliferation of CD4⁺T cell possibly so that tumor cells can escape immune surveillance and result in invasion and progression,thereby,contribute to the progression or metabasis of HCC,and can be used as an independent prognostic factor.

Objective To investigate the expression of HER4 in invasive breast cancer and its correlation with clinical characteristics and the prognosis.Methods Out of 58 patients with primary invasive breast carcinoma,32 underwent recurrence within 5 years and 26 did not.The expression of ER,HER2 and HER4 was detected by immunohistochemistry.The overall survival and relapse-free survival were followed up.Results The positive expression of ER,HER2 or HER4 was 31/58(53.4%),14/58(24.1%)and 19/58(32.8%),respectively.Non-recurrence group had a higher expression of HER4(P=0.05)and lower expression of HER2(P=0.043),but presented similar level of ER to that in recurrence group(P=0.866).Univariate analysis showed positive expression of HER4 had a significant relapse-free survival advantage(P=0.023)and a tendency of longer overall survival(P=0.057).However,HER2 manifested a negative effect on both relapse-free survival(P=0.048)and overall survival(P=0.02).In addition,coexpression status of HER2 and HER4 was aslo correlated with the relapse-free survival(P=0.016)and overall survival(P=0.014),with the best outcome in HER2-/HER4+ group.In multivariate analysis,more involved axillary lymphnodes,positive expression of HER2 and lack of HER4 expression were independently associated with a poor overall survival.The number of positive axillary lymphnodes and HER4 status were influencing factors for relapse-free survival as well.Conclusion HER4 status was an independent prognostic factor for invasive breast cancer and its positive expression predicts better outcome.

Objective To investigate the effects of all trans-retinoic acid(ATRA)on the expression of Akt or its phosphorylated proteins and on apoptosis of human gastric carcinoma cell line(SGC-7901).Methods The effects of ATRA on the cell proliferation was measured by methyl thiazolyl tetrazolium(MTT),the apoptosis of ATRA on SGC-7901cells was observed by fluorescentmicroscope,and the expression of Akt,p-Akt(Ser473)and p-Akt(Thr308)were measured by Western blot.Results ATRA inhibits the proliferation of SGC-7901 cells in a dose and time dependent manner.SGC-7901 cells showing typical apoptosis morphological changes were induced by ATRA at 48h.ATRA did not influence the expression of Akt,but significantly decreased the expression of p-Akt(Ser473)and p-Akt(Thr308).Conclution ATRA could induce apoptosis by down-regulating expression of p-Akt(Ser473)and p-Akt(Thr308).

Objective To explore the influence of RNA interference targeting Rab5a on invasion of breast cancer cells MCF-7.Method Four miRNA interference vectors targeting Rab5a were designed,constructed and transfected into breast cancer cell line MCF-7 each other.RT-PCR and Western blot were applied to selecte the most efficient group of miRNA interference suppression,then transwell assay were applied to analyse the cell invasiveness.Results RT-PCR analysis showed that cells MCF-7 transfected with Rab5a-miRNA-2 vector had the strongest inhibition of Rab5a gene,with the inhibition rate (50.01±2.69)%.Western blot analysis showed that cells transfected with Rab5a-miRNA-2 vector had the strongest inhibition of Rab5a protein,with the inhibition rate (68.49±3.70)%.Then we used Rab5a-miRNA-2 group for transwell assay.Transwell assay showed that the number of cells passing the membranes was 67.8±12.03 at 48h since transfecting,and was significantly different from the negative controls and blank controls(P＜0.01).Conclusion Rab5a silenced by miRNA expression vector can inhibit the invasiveness of breast cancer cells MCF-7.

Objective To study the prevention and treatment of complications of colostomy with carcinoma of large intestine.Methods 82 colostomies with carcinoma of large intestine were reviewed.The incidence and method of treatment of peristomal dermatitis,stomal necrosis,stomal hemorrhage,disassociation of skin and mucosa,infection,stomal retraction,stomal stenosis and prolapse of the stomal were analyzed.Results All stomal complications occurred in 24 patients(29.27%).The occurrence rates of partial necrosis,retraction and stenosis were higher in emergency operations than those in seleclive operations(χ²=8.2,P＜0.05).The complications of colostomy were mainly due to technical factors and could be prevented.The GOL(quality of life)of the patients with colostomy complications was depressed(t=3.325,P＜0.05).There were no death patients with the complications of colostomy in this group.Conclusions Sufficient preoperative intestinal preparation,rational chosing operative modality,location of colostomy and operative instruments properly selected,careful operative performation and postoperative treatment are the key points to avoid the complication and improve the life quality of the patients with carcinoma of large intestine.

Objective To investigate the effects of all trans-retinoic acid(ATRA)on the expression of MAPK signal transduction.Methods The effects of ATRA on the cell proliferation was measured by methyl thiazolyl tetrazolium(MTT),and the expression of ERK1/2,P38,JNK and their phosphorylated proteins were measured by Western blot.Results The inhibitory rate was 10.2%±0.5%,15.3%±0.5%,17.0%±0.7%,28.4%±1.0% and 36.9%±0.7%,respectively,as SGC-7901 cells were exposed to 10^-9mol/L-10^-5mol/L ATRA for 48 hours.ATRA did not influence the expression of ERK1/2,P38,JNK and p-JNK,but significantly decreased p-ERK1/2,significantly increased p-P38.Conclustion It is suggested that ATRA could inhibit proliferation of SGC-7901 cells via regulating the ERK1/2 and P38 MAPK pathway.

Objective To study the correlation between serum tumor marker and immumohistochemical marker of breast cancer.Methods To determine CA15-3 and CEA in serum of 65 breast cancer before surgery by electro chemiluminescent immunoassay(ECLIA).The expressions of C-erbB2,ER,PR in 65 breast cancer tissues were detected by immunohistochemistry,and analyze the correlation of them.Results CA15-3 was positively correlated with C-erbB2(r=0.597,P=0.000),negatively correlated but no significant with ER(r=-0.190,P=0.129),and PR(r=-0.179,P=0.153);CEA was positively correlated with C-erbB2(r=0.414,P=0.001),negatively correlated with ER(r=-0.267,P=0.031),negatively correlated but no significant with PR(r=-0.157,P=0.211);The positive rate of C-erbB2 in breast cancer was increased significantly with the clinical stage increasing(P=0.038),the positive rate of CA15-3 and CEA were increased with the clinical stage increasing(P=0.077,P=0.772).Conclusions CA15-3 was positively correlated with C-erbB2.CEA was positively correlated with C-erbB2,negatively correlated with ER;the positively rate of C-erbB2 in breast cancer was increased significantly with the clinical stage increasing,the positive rate of CA15-3 and CEA were increased with the clinical stage increasing.

Objective To investigate the relationship of chemokine receptor CXCR4 and vascular endothelial growth factor C(VEGF-C)with lymph node metastasis in breast carcinoma.Methods Immunohistochemistry was used to detect the expressions of CXCR4 and VEGF-C in 65 samples of breast carcinoma.The relationship between CXCR4,VEGF-C expressions and the clinicpathological parameters was statistically analyzed.Results The positive rate of CXCR4 expression with lymph node metastasis was higher than that without lymph node metastasis(57.9%vs 22.2%,P＜0.01).The positive rate of VEGF-C expression with lymph node metastasis was higher than that without lymph node metastasis(68.4%vs25.9%,P＜0.01).The positive expression of CXCR4 was consistent with the positive expression of VEGF-C,and the expression levels of CXCR4 and VEGF-C were closely correlated with lymph node metastasis,but not with age,tumor size,histological type,cellular differentiation and invasion depth.Conclusion CXCR4 and VEGF-C expressions may become predictors for the lymphatic metastasis of breast carcinoma.

Objective To investigate the expression of p53,ΔNp63 and ΔNp73 and their association with clinical pathology characteristics in different differentiated lung squamous cell carcinoma.Methods Expression of p53,ΔNp63 and ΔNp73 was detected by immunehistochemistry in 149 cases of different differentiated lung squamous cell carcinoma and 39 cases adjacent normal tissues.Results The positive rates of p53,ΔNp63 and ΔNp73 were 64.4%(96/149),85.9%(128/149),77.2%(115/149),respectively in lung squamous cell carcinoma.The positive rates and expression level in primary lung squamous cell carcinoma were remarkably higher than that in adjacent normal tissues(P ＜ 0.05).The expression of p53,ΔNp63 and ΔNp73 were not correlated with gender,age,clinical stage and metastasis in lymphoid nodes(P ＞ 0.05),however they were correlated with the degree of tumor differentiation(P ＜ 0.05).Conclusion p53,ΔNp63 and ΔNp73 is a group of valuable diagnostic markers of lung squamous cell carcinoma.p53,ΔNp63 and ΔNp73 may be as oncogenes to promote the progression of tumor.

Objective To explore and analyze imaging by bremsstrahlung phenomenon after ⁸⁹Sr administration in patients with bone metastases.And then monitoring distribution of the bone-seeking radiopharmaceutical ⁸⁹Sr in body and evaluating the feasibility of the method.Methods Bremsstrahlung imaging was performed in 15 patients in the 7th day after ⁸⁹Sr administration and there was comparision between it and ^99mTc-methylene bisphosphonate(MDP)bone scans.The region of interest(ROI)was outlined and then target/non-target ratio(T/NT ratio)was calculated.And a follow-up was made to observe thesis ache alleviation degree.Results Images gotten by ⁸⁹Sr bremsstrahlung imaging are similar to those by ^99mTc-MDP bone imaging,but the image reslution is less than the latter.T/NT ratio was about 3.13±1.03(1.24-5.81).Conclusion With skeletal metastases through calculating T/NT ratio,and with pain relief degree,⁸⁹Sr bremsstrahlung imaging for monitoring distribution of the bone-seeking radiopharmaceutical ⁸⁹Sr in body is feasibility.And it can preliminary decide prognosis of the patients.

Objective Owing to possible DNA breakage and degradation,tissues fixed by 10% neutral formalin may not be suitable for long-term preservation and downstream long DNA fragment amplification.This study aimed to compare the DNA quality extracted from lung cancer samples fixed by 70% ethanol versus neutral formalin.Methods Thirty cases of lung cancer tissues by 70% ethanol fixation(10 of which were collected by long time fixation more than 7 days)and 30 cases of formalin fixed samples were subject to DNA extraction with “Tissue/Cell DNA Extract kit”.Then DNA yields and purity were analyzed by agarose gel electrophoresis.Different length of expected DNA fragments of genes HBB(beta hemaglobin)and SERPINA9(antitrypsin)were amplified by common PCR and real-time quantitative PCR.Yields of PCR products were also compared by agarose gel electrophoresis.Results The integrity of DNAs extracted in 70% ethanol tissues was good without observed degradation,while DNAs from 10% formalin samples showed obvious smear bands during electrophoresis.Following common PCR,both yields of expected bands of 268bp and 536bp were more seen in group of 70% ethanol than in formalin group.By real time qPCR,all three amplicons of 100 bp,250bp and 350bp were more produced and shown at lower Ct values in 70% ethanol group compared with formalin group(P＜0.01).The success rate of qPCR for 346bp amplicons in formalin group samples was only 47%(14/30).DNA templates of antitrypsin were not significantly influenced by duration of ethanol fixation.Conclusion Quality of DNAs from 70% ethanol-fixed specimens was superior to formalin-fixed samples and suitable for long fragment amplification.Ethanol-fixed samples may be also suitable for long time transportation without compromising the DNA quality.

Objective To investigate the effect of combined detection of serum tumor markers,carcinoembryonic antigen(CEA),carbohydrate antigen 125(CA125),cytokeratin fragment 19(CYFRA21-1),neuron-specific enolase(NSE),and squamous cell carcinoma antigen(SCC),in the clinical diagnosis of lung cancer.Methods 144 blood samples of the subjects were collected,including 48 adenocarcinomas,33 squamous carcinomas and 31 small cell cacinomas of lung cancer patients;32 blood samples of normal persons as control.All the diagnosis were supported by histology.The serum level of CEA,CA-125,CYFRA21-1and NSE were detected by electrochemistry luminescent technique and SCC by microparticle enzyme immunoassay(MEIA)technology.Sensitivities,specificities and positive predictive values were calculated.Results Sensitivities of CEA,CA-125,CYFRA21-1,NSE,and SCC were 54.46%,67.86%,39.29%,38.39%,and 28.57% respectively,while specificities were 93.75%,96.88%,93.75%,87.50%,and 96.88% respectively.The combined detection of positive predictive value of total serum tumor markers was 92.17%,sensitivity and specificity were 94.64% and 71.87%.Conclusion The combined detection of CEA,CA-125,CYFRA21-1,NSE,and SCC is an effective method in clinical diagnosis of lung cancer that could increase the sensitivity of the diagnosis of lung cancer and the detection rate.

Objective To evaluate the efficacy and safety of gemcitabine and cisplatin versus navelbine and cisplatin in anthracycline-and(or)taxane-resistant metastatic breast cancer(MBC).Method A total of 68 patients with metastatic breast cancer were randomly assigned to receive the regimen of GP(n=36)or NP(n=32).Group GP:Gemcitabine 1000mg/m²,d1,d8;DDP 25mg/m²,d1-3.Group NP:Navelbine 25mg/m2,d1,d8;DDP25mg/m²,d1-3.Two groups were treated at least 2 courses with 3 weeks interval,efficacy was evaluated after 2 courses of chemotherapy.Results The response rate was 55.6%(20/36)in group GP,and 53.1%(17/32)in group NP.The degree III-IV thrombocytopenia was more common in group GP than in the group NP,but the degree phlebitis was more serious in group NP.Conclusion NP and GP for treating refractory metastatic breast have a high response rate,there was no significant difference between two groups(χ²=0.0403,P=0.84).Two groups have tolerable side effects.

Objective The analysis made by comparing the Ultrasound characteristics with pathologic result of thyroid tumor,improve coincidence rate of diagnosis by Color Doppler Ultrasound.Methods The paper reviewed 150 patients of thyroid tumor that have been confirmed by postoperative pathology,including of 37 thyroid adenomas,78 nodular goiters,35 thyroid carcinoma.Analysis made by comparing the 2D and the color doppler ultrasound characteristics with pathologic examination.Results The interior and ambitus Vmax and RI of thyroid carcinoma are obviously highly thyroid adenomas and nodular goiters.The specificity of diagnosis is highly,such as peplos and circularity blood of the thyroid adenomas;low echo-level and calcification of thyroid carcinoma.But thyroid adenomas and nodular goiters easily misdiagnosis because of diverseness tumor.Conclusion Color doppler ultrasound may diagnose the thyroid tumor properties(substantive or cystic mass、mixed),the benign and malignancy,observing blood b-flow conditions and the internal distribution of operation treatment,has certain directive significance,and it is a simple,inexpensive,accurate and good repeatability,inspection method for routine inspection of thyroid preoperative.

Objective To observe the effect of concurrent chemoradiotherapy plus adjuvant chemotherapy in patients with advanced cervical squamous cell carcinoma.Method 156 cases with stage Ⅱa-Ⅲb cervical squamous cell carcinoma were randomly divided into both the concurrent chemoradiotherapy group(experimental group)and radiotherapy group(control group).Radiotherapy was administered by intracavity and external beam radiation therapy,dose of point A were given for 40Gy-48Gy/10f-12f and dose of point B for 46Gy-50Gy/23f-25f.In the same time,experimental group was treated by DDP 40 mg/d 1,3,5 intravenous drip,repeated every week.Tenth day after radiation therapy,adjuvant chemotherapy using TP program was given three times,21 days intervally.Results The short-term efficiency for the experimental group and the control group was 88.61% and 75.33% respectively (P＜0.05).In one year to three year after treatment,the overall survival rate(OS)and distance metastasis free survival rate(DMF)and local recurrence free(LRF)for the experimental group were different significantly (P＜0.05)than the control group.Quality of life of were significantly improved after treatment.Lack of appetite,diarrhea and blood toxicity of the treatment modalities were observed as limiting toxicity(P＜0.05).Conclusion Concurrent chemoradiotherapy plus adjuvant chemotherapy for advanced cervical cancer patients can improve short-term and quality of life,the limiting toxicity can be tolerated.