Purpose To investigate the expression of EZH2 mRNA and protein, and its relationship with tumor cell proliferation in human prostate carcinoma (PCa).Methods A tissue microarray was constructed, which contained 68 dots of formalin-fixed,paraffin-embedded tissue samples, including 48 cases of human PCa.Immunohistochemical markers, including EZH2 and Ki-67,were used on the tissue microarray sections by the immunohistochemical staining method.In situ hybridization (ISH) using an EZH2 oligonucleotide probe was also performed on the tissue microarray sections.Additional 15 cases of benign prostate hyperplasia (BPH) and 12 cases of high grade prostate intraepithelial neoplasia (HGPIN) were used as controls.Results The positive rates of EZH2 protein and mRNA expression were 87.50% and 81.25% in the PCa, 16.67% and 16.67% in the HGPIN,13.33% and 6.67% in the BPH,respectively. There were statistical difference between PCa, HGPIN and BPH, respectively (P<0.05).The expression of EZH2 protein and mRNA had no statistical difference (P>0.05).The expression of EZH2 protein was related to Gleason score, TNM stage (P<0.05), but not to age and serum prostate-specific antigen (PSA) level (P>0.05). The expression of EZH2 mRNA was related to TNM stage (P<0.05),but not to age, PSA and Gleason score (P>0.05).The staining intensity of EZH2 was positively correlated with the Ki-67 indexes (r=0.746,P<0.05).Conclusions Over-expression of EZH2 which is involved in accelerating proliferation may play an important role in the pathogenesis and progression of human prostate carcinoma. As a molecular marker of prostate carcinoma, EZH2 may serve as a new index for estimating the level of malignancy and progression of the prostate cancer.

Objective To study the effects of midkine (MK) gene-targeting small interfering RNA (siRNA)on the invasion of melanoma cells.Methods Three MK gene-targeting siRNAs (S1,S2 and S3)were designed,constructed,and transfected into human A375 melanoma cells.Real-time PCR was performed to measure the expression of MK gene and to screen the siRNA with best efficacy.Then,A375 cells were transfected with the optimal siRNA of various doses (3.125,6.25 and 12.5 nmol/L)followed by additional culture of various durations(24,48,72 hours).Some A375 cells remaining untreated served as the blank control group,and some transfected only with liposomes served as the vector control group.Reverse transcription (RT) -PCR and Western blot were conducted to detect the mRNA and protein expression of MK,respectively,MTT assay to observe the adhesion of A375 cells,and Boyden chamber was used to evaluate cell invasion.Results The expression of MK mRNA was downregulated by all the three siRNAs,especially by the siRNA S3,which was used in the following transfection experiment.Real-time quantitative PCR revealed that the MK mRNA expression was reduced by the siRNA in a dose- (r24hours=-0.906,r4Bhours=-0.922,r72hours=-0.939,all P<0.01)and time-dependent(r3.125nmol/L=-0.889,r625nmol/L=-0.935,r125nmol/L=-0.928,all P<0.01)manner.MTT assay showed that the percentage of adhesing cells was 73.66%±2.25%,49.36%±2.16%and 28.35%±1.68%in A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively.The number of cells migrating across the chamber filter was 23.9±1.6,12.1±1.5,5.6±1.2 among A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively,significantly lower than that in the blank control group(36.8±1.5).The percentage of adhesing cells and number of migrating cells decreased with the dose of siRNA(r=-0.936,-0.915,P<0.01,0.05,respectively).Conclusions MK gene might play an important role in the adhesion and invasion of melanonla cells.To down-regulate the expression of MK gene by siRNA may suppress the adhesion and invasion of melanoma cells.

Objective To evaluate the causes and prognosis of severe trauma in the elderly.Methods The 168 patients in elderly group (aged 60 to 91 years), 517 in middle-aged group (aged 36to 59 years) and 405 in young group (aged 18 to 35 years) were evaluated using an abbreviated injury scale (AIS2005) and injury severity score (ISS). All patients with ISS ≥ 16 were selected during a seven-year period. The injury severity, injury site number, cause of injury, injury site, emergency operation, diseases before injury, secondary infection after injury, development of multiple organ dysfunction, number of patients with Intensive Care Unit (ICU) stay, length of stay in ICU and prognosis were compared among three groups. Results The main cause of injury was accident (64patients, 38.1%), followed by traffic accident (63 patients, 37.5%) in elderly group. The traffic accident was major cause of injury in middle-aged and young group (246 patients, 47.6%; 153patients, 37.8%, respectively), followed by fall from high places (128 patients, 24.8%; 102 patients, 25.2%, respectively). The main injury sites were head and chest in elderly, middle-aged and young group (155 patients, 92.3%; 411 patients, 79.5%; 321 patients, 79.3%, respectively).There were significant differences among three groups in injury site number, emergency operation,pre-injury diseases, secondary infection after injury, number of patients with ICU stay and length of stay in ICU (F=8. 299, P＜0.01; x2= 14.88, P=0.001; x2=254.6, P＜0.01; x2=10. 54, P=0. 005; x2 = 15.62, P＜0.01; F= 5.760, P= 0.005, respectively ). In spite of injury severity (F=2.950, P= 0.053), there were significant differences between elderly group and middle-aged or young group (t=2.325, P=0.021; t=2.128, P=0.034, respectively). The incidence of multiple organ dysfunction had no significant difference among the three groups (x2 = 1.142, P= 0.565). The cure rate and unhealed automatically discharged patients had significant differences (x2 = 13.77, P= 0. 001;x2 =6.025, P= 0.049, respectively). The mortalities were similar (x2 = 1.397, P= 0.497). The leading cause of death among three groups was a serious head injury. Conclusions For elderly patients, it is important to reduce accidental injuries and traffic accidents, to improve the cure rate,and to reduce the unhealed and mortality rate.

OBJECTIVE To observe the expres-sion of interleukin-12 and eosinophils in the nasal mu-cosa of allergic rhinitis mice. METHODS Thirty nine male BALB/c mice,6～8 weeks old,were randomly divided into three groups: control group,allergic rhinitis (AR)group, and Budesonide treatment group. Al-lergic rhinitis model in mice were established by using ovalbumin intraperitoneal immunization and nasal anti-gen challenge. The nasal mucosa obtained from mice of three groups were stained routinely by HE and im-munohistochemical method to observe the distribu-tion and expression of interleukin-12 and eosinophils. RESULTS The expression of eosinophils in the nasal mucosa of AR group was significantly higher than con-trol group(P

Objective To screen for and validate unknown tumor suppressor genes (TSGs) in sporadic colorectal cancer (CRC) patients.Methods Through loss of heterozygosity (LOH) analysis on chromosome 10 in sporadic CRC,we have found D10S185 (10q23.31-24.33 ) exhibit a higher LOH frequency in our previous study.In present study,we screen for unknown TSGs in this region through the microarray.The expression of the new gene was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR).RT-PCR,immunohistochemistry and Western blot were done in colorectal cancer tissues with their pair-matched normal tissues in 50 cases to validate the results of microarray.Results Through the microarray-based high throughput screening,we found 4 significant down-regulated genes:PLCE1,CPEB3,NKX2-3 and SEMA4G,among them the down-regulation of PLCE1 was most significant.The results of qRTPCR were in relative agreement with the DNA microarray data.RT-PCR,immunohistochemistry and Western blot also showed that the expression of PLCE1 was at low levels in 46％ cancer tissues compared with normal tissues,more frequent in the poor differentiation tumor in patients under age 60 years (P ＜ 0.05 ).Conclusions This study demonstrated that down-regulation of PLCE1 was related to the tumorigenesis of sporadic colorectal cancer.PLCE1 might play a suppressive role in the development of colorectal cancer.

ObjectiveTo study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA(siRNA) on adhesion and invasion of human breast cancer cell. MethodsReal time PCR was used to evaluate the TROP-2 mRNA of seven human breast cancer cell lines Bcap-37 ,LCC1 ,MCF-7 ,MDA-MB-231,MDA-MB-435, MDA-MB-468 ,and ZR75-1. The cell line of TROP-2 highest expression was transfected with different dose of TROP-2 siRNA. The expression of TROP-2 mRNA and protein were determined by Real-time quantitative PCR and immumoflurescence method. The cell adhesion was evaluated by MTT assay,and invasion was exmined by hoyden chamber,respectively. Results Cell line MCF-7 showed the highest elevation of TROP-2 mRNA in seven breast cancer cell lines. The results from real-time quantitative PCR and immumoflurescence method showed that TROP-2 mRNA and protein reduced in time-and dose-dependent manners( P ＜ 0.01 ;P ＜ 0.01 ). The adhesive rate of siRNA groups(5 nM,10 nM,and 20 nM)was(52.9 +2.5)％ ,(25.6 ±2.3)％, ( 12.8 +2.2)％ (P ＜0.01 ) ,respectively.The transwell results showed that the invasion cells was(78 ± 17), (39 ± 15), ( 19 ± 16), ( 136 +25 ) and( 139 ±21 )in different groups(5,10,20 nM siRNA,and controls) ,respectively(P ＜0.01). ConclusionTROP-2 gene might play an important role in adhesion and invasion of human breast cancer cell. siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cell.

Objective To explore the diagnosis and treatment of primary small bowel tumors. Methods The clinical data of 58 cases of primary small bowel tumors were retrospectively analyzed. Results Fifty-eight cases of primary small intestinal tumors were comfirmed by operation and/or pathology .Of them,19 suffered from benign tumors and 39 from malignant tumors.The main clinical manifestations of small bowel tumors were abdominal pain, abdominal mass, ileu and upper gastrointestinal hemorrhage.Twenty cases were diagnosed before the operation,and the misdiagnosis rate was 65.5%.All the 19 benign small intestinal tumors underwent local intestinal resection.In 39 malignant cases,18 underwent radical excision of the tumors, 6 received palliative excision and 5 underwent intestinal bypass operation. One patient died after the operation.The 1,3,5 -year survival rate of malignant tumors was 57.1%,28.6%and 9.5% respectively. Conclusions Primary small bowel tumor is uncommon and easy to be misdiagnosed.So attention must be paid to this disease.Endoscopy, X-ray ,BUS, CT and capsule endoscopy are the ideal diagnostic methods for small intestinal tumors .

Objective To evaluate salvage liver transplantation(LT)for postoperative tumor recurrence or liver dysfunction in patients of hepatocellular carcinoma(HCC).Methods From April 2001 to March 2006,97 HCC patients with the tumor within Milan Criteria underwent LT as the primary treatment(71 cases)and salvage LT because of after resection tumor recurrence(n=20)or postoperative liver dysfunction(n=6).Perioperative and postoperative parameters and long-term survival were compared between the groups of primary LT and salvage LT.Results The mean age(50.0 years vs.49.7 years),gender,and etiology of liver disease(hepatitis B/C/nonviral)were comparable between the two groups.In the salvage LT group.the mean time between liver resection and LT was 2.50 years.Clinical characteristics such as tumor number(1.37 vs.1.50),operative time(7.92 hours vs.8.56 hours),blood loss (1981.69 ml vs.2626.92 ml)and transfusion(1981.69 ml vs.2626.92 ml)were not statistically different (P＞0.05)between the two groups.The size of largest tumor was significantly different between salvage LT group and the primary LT group(2.81 cm vs.2.05 cm)(t=2.298,P=0.028).By a median follow up of 14.63 months,overall survival after liver transplantation was not different between the 2 groups(X2=0.003.P=0.959).Conclusion In selected patients,liver resection prior to transplantation does not increase the morbidity or impair long.term survival following LT.Therefore.1iver resection prior to transplantation can be integrated with the treatment strategy for HCC.

Objective To investigate the diagnosis and treatment of hepatic metastasis from gastrointestinal stromal turnor(GIST).Methods The clinical data of 16 patients with GIST who had been admitted to our hospitalfrom December 1993 to May 2007 were retrospectively analyzed.Results Of all patients,14 underwent radical resection and 2 underwent palliative operation.Two patients with palliative operation and 3 with radical resection were administered with imatinib postoperatively. All patients were followed up for 3-161 months,and GIST metastasis and invasion was observed in 8 of the 14 patients who received radical resection.Of the 7 patients with hepatic metastasis.3 were treated with hepatic artery chemoembolization,1 was administered with imatinib,2 received reoperation and 1 did not receive any treatment. Reoperation was carried out on 1 patient who had abdominal wall metastasis.The 1-and 3-year survival rates of the 16 patients were 92%and 74%,respectively.Conclusions The recurrence rate of GIST after hepatectomy is high.Complete surgical resection is the best curative treatment for hepatic metastasis from GIST and GIST recurrence.The combination of surgical resection and imatinib administration may help to improve the prognosis of patients with hepatic metastasis from GIST.

Objective To investigate the effects of preventive chemotherapy for hepatocellular carcinoma (HCC) exceeding Milan criteria after liver transplantation. Methods The clinical data of 243 patients who had undergone orthotopic liver transplantation for HCC exceeding Milan criteria from April 2001 to July 2007 were retrospectively analyzed. Of all patients, 162 received preventive chemotherapy after transplantation. Results The 1- and 3-year survival rates and disease-free survival rates were not statistically different between patients who had received chemotherapy (78.5%, 63.7% ; 76.8%, 52.5% ) and those without chemotherapy (56.6%, 39.1%; 69.3%, 64.7% ) (X2 = 3.084, 0.444, P > 0.05). Cox regression analysis demonstrated that postoperative chemotherapy was not an independent factor affecting the survival rates of HCC patients without vascular invasion, but an independent factor affecting the survival rates of HCC patients with vascular invasion. Conclusions Early preventive chemotherapy could obviously increase the survival rate and delay the tumor recurrence of patients with HCC exceeding Milan criteria, especially for HCC patients with vascular invasion.