Objective To investigate the expressions of Hedgehog sisnaling pathway genes in hepatocellular carcinoma tissues(HCC),and the effect of specific Hedgehog pathway inhibitor(KAADcyclopamine)on the growth of HCC cells and the expressions of Hedgehog genes. Methods The expression of Hedgehog signaling pathway components(Ihh,Ptch,Smo and Gli)was investigated in 14 HCC tissue slices,4 HCC cell lines and a normal hepatic cell line by using immunochemistry.The expression of Ihh,Ptch,Smo and Gli proteins was investigated in 9 HCC tissue specimens and 6 normal hepatic tissue specimens by using Western blotting.The expression of Ihh、Ptch、Smo、Gli and Hip genes was investigated by RT-PCR.Results The positive ratio of Gli,Ptch,Ihh and Smo were 42.9%,71.4%,71.4% and 85.7% in 14 HCC tissue slices,respectively.The expressions of Gli protein and Gii gene were up regulated while the expression of Hip gene was down regulated in HCC specimens compared with normal hepatic tissue specimens.Hedgehog signaling pathways in HCC cell lines HepG2,Bel-7402 and QGY-7701 were activated;KAAD-cyclopamine,a specific inhibitor of the Hedgehog signaling pathway,down regulated cell growth and the expressions of Ptch and Gli genes in the 3 HCC cell lines(Ptch gene:tHepG2=3.78,tBel-7402=9.03,tQGY-7701=5.63;Gli gene:tHepG2=9.61,tBel-7402=4.15,tQGY-7701=20.30,P＜0.05 in each group).The expression of Hip gene was up regulated in QGY-7701 after treated with KAAD-cyclopamine(t=4.70,P＜0.05).Conclusion The expression of main Hedgehog signaling pathway components were detected in HCC,KAAD-cyclopamine specifically inhibited the Hedgehog signaling pathway.

Objective: To determine the relationships between apoptosis induced by transforming growth factor beta 1 (TGF-β1) and Smad in human hepatoma cell lines. Methods: Three human hepatic carcinoma cell lines, involving different status of the p53 gene respectively, were used in this study.TGF-β1-induced apoptosis in hepatic carcinoma cell lines was quantitated using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. For identification of the mechanism of apoptosis induced by TGF-β1, these cell lines were transfected with a TGF-β1-inducible luciferase reporter plasmid containing Smad binding elements (SBE) and luciferase gene using LF2000, then were treated with TGF-β1. Relative luciferase activity was assayed respectively. Results: Among three cell lines studied with TUNEL assay, addition of TGF-β1 induced apoptosis only in HepG2 cells (wild type p53). In contrast, Huh-7 ( mutant p53) and Hep3B ( deleted p53) cell lines lacked apoptosis. The detection of luciferase activity indicated that HepG2 cells dramatically increased the response to TGF-β1 induction, Huh-7 and Hep3B cell lines significantly lowered luciferase expression. Conclusion: HepG2cells were highly susceptible to TGF-β1-induced apoptosis compared with Hep3B and Huh-7 cell lines.Smad4 may be a central mediator of the TGF-β1 signaling transdution pathway.

Objective To investigate factors affecting the metastasis of lymph nodes around the root of inferior mesenteric artery(IMA) in rectal cancer,and the significance of root lymph nodes dissection of IMA in radical surgery for rectal cancer.Methods Clinicopathological data of 105 rectal cancer patients undergoing root lymph node dissection of IMA during radical resection in Peking University First Hospital from January 2005 to December 2008 were analyzed retrospectively.Rectal cancer patients without root lymph node dissection of IMA during the same period served as control.Results were compared between these two groups for survival and local recurrence rates.Results The rate of lymph node metastasis around the origin of IMA was 9.5％ (10/105).The five-year survival rate in patients with IMA root nodal dissection was 71.3％,and that without was 70.6％ (P =0.995),while the local recurrence was respectively 1.9％ and 7.4％ (P ＜ 0.05).In multivariate analyses,IMA root nodal metastasis occurred more frequently in patients with pT3 and pT4 tumor(Wald =5.764,P ＜ 0.05) and poorly differentiated tumor(Wald =7.818,P ＜ 0.05).Conclusions Root lymph nodes dissection of IMA could not increase five-year survival rate,but it could reduce local recurrence rate in patients with rectal cancer.In radical surgery of rectal cancer,lymphadenectomy of IMA root should be performed in patients with T3 and T4 tumor with poorly differentiated tumor,so as to reduce local recurrence rate.

Objective To study the presentation,sensitivity of different diagnostic methods,snrgical modalities and pathological findings for patients of substemal goiter. Methods The clinical data of fifty nine cases of substernal goiter surgically treated were retrospectively analyzed. Results The main symptoms of substernal goiter patients were asymptomatic cervical mass(39/59),airway obstruction (13/59),hyperthyroidism(4/59),hoarseness(3/59)and choke(3/59).The sensitivity of chest X-ray,ultrasonography,CT and scintigraphy was 62.8%,15.8%,85.7%,and 50.0%respectively,and the specificity Was 99.4%,99.8%,99.5%and 99.0%,respectively.Standard cervical approach was successful for thyroidectomy operations in 57 cases.One patient underwent thyroidectomy by sternotomic approach,and another one by combined cervici-sternotomic approach.No major complications or perioperative deaths occurred.Pathology revealed nodular goiter in 48 cases.Grave's disease in one,thyroid adenoma in 2,and carcinoma in 8. Conclusions CT and chest X-ray are sensitive diagnostic techniques for substerual goiter.Cervical approach is appropriate for most substernal goiter.

OBJECTIVE To investigate the mechanisms for carbapenem resistance in clinical isolates of Pseudomonas aeruginosa from surgical intensive care unit(SICU).METHODS The minimal inhibitory concentrations(MICs) of various antibiotics were detected with agar dilution method;their ?-lactamases were extracted,types of their enzymes were identified and then their physical and chemical characteristics were studied by three dimentional extract test and isoelectric focusing(IEF) electropheresis;primers specific to IMP and VIM genes were utilized to amplify the metallo-?-lactamase genes.The levels of outer membrane protein OprD2 were measured by Western blot.RESULTS Forty one strains were resistant to imipenem from 49 strains of P.aeruginosa isolated from SICU during 3 years.Among them,34 isolates were found to produce high level of AmpC enzymes.Eight produced extended-spectrum ?-lactamases(ESBLs) at the same time.2 produced ESBLs only.No metallo-?-lactamases were detected.The activity of enzymes in the imipenem resistant group(74.32?53.42) was statistically different from the sensitive one(8.7?16.16,P

Objective To evaluate clinical risk factors for Meropenem resistant Pseudomonas aeruginosa (MRPA) infection in surgical intensive care unit. Methods Retrospective case-control study was designed. Resultant MRPA and Meropenem sensitive Pseudomonas aeruginosa ( MSPA) cases were paired with respective control group. Candidate clinical factors including gender, malignancy, duration of ICU stay, length of operation, volume of blood loss, transfusion, kind of incision ( clean/contaminated/ infected) , and previous antibiotics history were examined by bivariates and multivariate regression. Results Risk factors for MRPA infection included duration of ICU stay before infection (odds ratio [ OR] = 2. 234, 95% confidence interval [ CI] = 1. 710 - 2. 918), administration of third generation cephalosporins (OR = 2. 885, 95% CI = 1. 133 - 7. 345) and Imipenem ( OR = 1. 238, 95% CI = 1. 469 - 35. 659). A long ICU stay (OR = 2. 284, 95% CI = 1. 772 - 2. 945), malignancy (OR =4.796, 95%CI = 2.001 -11.491), previous medication of quinolones ( OR = 3.563, 95% CI = 1.499 - 8.468) and third generation cephalosporins (OR = 3. 796, 95% CI = 1. 543 - 9. 343) were associated with MSPA infection. Conclusion MRPA infection was affected by multiple clinical factors. Short ICU stay, restrict use of third generation cephalosporins and Imipenem may help to prevent MRPA infection.

Objective To investigate the surgical treatment strategies and prognostic factors of cT4bM0 primary rectal cancer.Methods The clinical data of 53 patients with cT4bM0 primary rectal cancer who were admitted to the First Hospital of Peking University from January 2000 to December 2010 were retrospectively analyzed.All the patients received en-bloc multivisceral resection and postoperative chemotherapy.The survival and prognostic factors were analyzed.The patients were followed up via out-patient examination,phone call or mail,and the follow-up was ended till December 2012.The survival curve was drawn using the Kaplan-Meier method,and the survival was analyzed using the Log-rank test.Uni-and multivariate analysis were done using chisquare test and COX's proportional hazard model.Results Of all the 53 patients,20 received posterior pelvic exenteration (PPE),20 received total pelvic exenteration (TPE),3 received low anterior resection (LAR) +local resection of ballder,2 received LAR + ovariectomy,2 received LAR + local resection of ureter,1 received LAR + local resection of posterior vaginal wall,1 received LAR + vesiculectomy and vesectomy,3 received abdominoperineal resection (APR) + local resection of posterior vaginal wall,1 received APR + sacrectomy.R0 resection was achieved in all the patients.No intraoperative death was observed,and the incidence of postoperative complication was 9.4％ (5/53).The results of postoperative pathological examination showed that 2 patients were with well-differentiated adenocarcinoma,41 with moderate-differentiated adenocarcinoma,and 10 with poorly differentiated adenocarcinoma.Twenty-four patients were with lymph node metastasis.Four patients were in TNM stage Ⅰ,25 in TNM stage Ⅱ and 24 in TNM stage Ⅲ.Fifty-three patients were followed up postoperatively,and the median time for follow-up was 33 months (range,4-116 months).The overall 5-year survival rates was 57.3％.The 5-year survival rate for patients with or without lymph node metastasis were 77.1％ and 30.4％,respectively,with significant difference between the 2 groups (x2 =7.374,P ＜ 0.05).The 5-year survival rates of patients with malignant infiltration and inflammatory adhesion mere 51.0％ and 68.5％,with no significant difference (x2=1.148,P ＞0.05).The 5-year survival rates of patients with malignant infiltration and inflammatory adhesion in stage Ⅱ were 74.6％ and 85.7％,with no significant difference between the 2 groups (x2=0.118,P ＞ 0.05).The 5-year survival rates of patients with malignant infiltration and inflammatory adhesion in stage Ⅲ were 28.8％ and 37.5％,with no significant difference between the 2 groups (x2 =0.959,P ＞ 0.05).The results of univariate analysis showed that lymph node metastasis and TNM stage were the risk factors influencing the prognosis of patients with cT4bM0 primary rectal cancer (x2=6.468,6.596,P ＜ 0.05).The results of multivariate analysis showed that lymph node metastasis was the independent risk factor (RR =3.797,P ＜ 0.05).Conclusions En-bloc multivisceral resection should be the first surgical treatment choice for patients with cT4bM0 primary rectal cancer,and lymph node metastasis is the independent risk factor.Under the same N stage,the prognosis of patients with malignant infiltration or inflammatory adhesion is similar if R0 resection is achieved.

ObjectiveTo explore the clinical diagnosis and surgical treatment of primary retroperitonealneurilemoma(schwannoma). MethodsClinicaldataof 47patientsof primary retroperitoneal schwannoma admitted and surgically treated from January 1995 to December 2009 were retrospectivelly reviewed.ResultsAs diagnosed by pathology there were 36 cases of Benign schwannoma,with a median age at onset of 41years, among those 11 patients were symptomatic, and 25 were asymptomatic. There were 11 malignant 11 cases, the median age was 38 years, among those 6 patients were symptomatic, and 5 were asymptomatic. The positive diagnostic rate of preoperative CT and MRI were 36. 2％ ( 17/47 ) and 58. 3％ ( 7/12 ) respectively. Immunohistochemically positive rates of S-100 were 100％ and 81.8％(9/11) in benign and malignant group respectively.All cases underwent surgical treatment. Surgical resection rates for benign and malignant groups were 100％ and 90. 9％(10/11)respectively. There was no perioperative death, Overall 5-year survival rates were 100％ and 45.5％ for benign and malignant tumors groups respectively. In benign group 2 cases recurred, in malignant group 4 cases recurred, and 3 had distant metastasis.ConclusionsPrimary retroperitoneal schwannomas are less common. It is difficult to make an accurate preoperative diagnosis. Surgery is the most effective therapy.Prognosis is good for benign and poor for malignant retroperitoneal neurilemomas.

Objective To investigate the correlation between VEGF-C and COX-2 expression in human rectal cancers and its significance in cancer metastasis. Methods VEGF-C expression was detected with Western blot in LOVO cells treated with NS-398 or PGE2. VEGF-C and COX-2 expression in 45 rectal adenocarcinomas was tested with immunohistochemistry. Results NS-398 inhibited the VEGF-C expression, and PGE2 up-regulated the expression of VEGF-C in a dosage-dependent way in LOVO cells. VEGF-C expression was significantly higher in adenocarcinomas with lymph node metastasis, and was related with the expression of COX-2 in 45 rectal adenocarcinomas. Conclusion COX-2 up-regulates VEGF-C and VEGF-C plays an important role in lymphatic metastasis of rectal cancers.

Objective To evaluate total pelvic exenteration (TPE) in the treatment of locally recurrent rectal cancer (LRRC). Methods Clinical data of 35 patients with LRRC who underwent TPE between 1989 and 2003 were analyzed retrospectively. Results Thirty patients underwent TPE, the remaining 2 did sphincter-preserving TPE, 2 with lower sacrectomy and 1 with hemipelvectomy, among them 80% cases received radical resection. Operative mortality rate was 3%, and morbidity rate was 51%. The overall post TPE tumor local recurrence rate was 48%. The 5-year survival rate was 16% in all cases and 19% in radical resection group. The 5-year survival rate in patients without lymph node metastasis was 24%, and 0 in patients with metastasis. Conclusion Effective TPE treatment lies in strict patient selection and radical resection.