Neuropilin-1 is up-regulated by cancer-associated fibroblast-secreted IL-8 and associated with cell proliferation of gallbladder cancer.

We previously demonstrated that cancer-associated fibroblasts (CAFs) promoted the proliferation of gallbladder cancer (GBC) cells, but the mechanism is not clear. Neuropilin-1 (NRP-1) plays an important role in various malignancies as transmembrane glycoprotein. Our goal was to reveal the relationship between CAFs and NRP-1 and their potential functions in GBC. In this study, we found NRP-1 was overexpressed in GBC tissue, associated with poor survival and was up-regulated by CAFs. The cytokine array cluster analysis revealed IL-8 secreted by CAFs facilitated the up-regulation of NRP-1 in tumour cells. NRP-1 knockdown suppressed tumour growth in vivo. Gene expression microarray analysis showed 581 differentially regulated genes under NRP-1 knockdown conditions. Ingenuity pathway analysis demonstrated that NRP-1 knockdown may inhibit tumour progression by affecting cell proliferation. We then confirmed that NRP-1 knockdown in NOZ and GBC-SD cells significantly inhibited cell proliferation. Additionally, the IL-8 mediated MDM2 and CCNA2 expression were affected by NRP-1 knockdown. Our findings suggested that NRP-1 was up-regulated by CAF-secreted IL-8, which subsequently promoted GBC cell proliferation, and these molecules may serve as useful prognostic biomarkers and therapeutic targets for GBC.

Optimal number of harvested lymph nodes for curatively resected gallbladder adenocarcinoma based on a Bayesian network model.

BACKGROUND AND OBJECTIVES: To identify the optimal range and the minimum number of lymph nodes (LNs) to be examined to maximize survival time of patients with curatively resected gallbladder adenocarcinoma (GBAC). METHODS: Data were collected from the surveillance, epidemiology, and end results database on patients with GBAC who underwent curative resection between 2004 and 2015. A Bayesian network (BN) model was constructed to identify the optimal range of harvested LNs. Model accuracy was evaluated using the confusion matrix and receiver operating characteristic (ROC) curve. RESULTS: A total of 1268 patients were enrolled in this study. Accuracy of the BN model was 72.82%, and the area under the curve of the ROC for the testing dataset was 78.49%. We found that at least seven LNs should be harvested to maximize survival time, and that the optimal count of harvested LNs was in the range of 7 to 10 overall, with an optimal range of 10 to 11 for N+ patients, 7 to 10 for stage T1-T2 patients, and 7 to 11 for stage T3-T4 patients. CONCLUSIONS: According to a BN model, at least seven LNs should be retrieved for GBAC with curative resection, with an overall optimal range of 7 to 10 harvested LNs.

Decision of surgical approach for advanced gallbladder adenocarcinoma based on a Bayesian network.

BACKGROUND AND OBJECTIVES: To determine whether radical resection can benefit patients with advanced gallbladder adenocarcinoma using a Bayesian network (BN) with clinical data. METHODS: In total, 362 patients who had undergone surgical treatment of gallbladder adenocarcinoma at a tertiary institute were evaluated to establish two BN models using a tree-augmented naïve Bayes algorithm. We then chose 250 patients with T3-4N0-2M0 stage gallbladder adenocarcinoma to test the posterior probability after the surgical type was taken into account. RESULTS: In total, 170 patients (≤7 months) and 137 patients (>7 months) were correctly classified in the median survival time model (accuracy, 84.81%), and 204 patients (≤12 months), 15 patients (12-36 months), 17 patients (36-60 months), and 34 patients (>60 months) were correctly classified in the 1-, 3-, and 5-year survival model (accuracy, 74.59%), respectively. Every posterior probability in the two models upregulated the ratio of the longer survival time and suggested a better prognosis for gallbladder adenocarcinoma that can be improved by R0 resection. CONCLUSIONS: These BN models indicate that stages T4 and N2 gallbladder adenocarcinoma are not contraindications for surgery and that R0 resection can improve survival in patients with advanced gallbladder adenocarcinoma.

Squamous/adenosquamous carcinoma of the gallbladder: Analysis of 34 cases and comparison of clinicopathologic features and surgical outcomes with adenocarcinoma.

OBJECTIVES: To explore clinicopathological features and effects of surgical treatment of squamous/adenosquamous carcinoma of the gallbladder. METHODS: We enrolled 411 patients who were surgically treated for gallbladder cancer in our hospital, including 10 with squamous cell carcinoma (SCC), 24 with adenosquamous carcinoma (ASC), and 377 with adenocarcinoma (AC). The ASC-SCC group was compared with the AC group for clinicopathological features and surgical outcomes. RESULTS: The patients' average age was 61.4 years. Abdominal pain was the most common presenting symptom, and 67.6% of patients had gallstones. All patients had advanced-stage (T3/T4) carcinomas. The ASC-SCC group had significantly higher percentages of T4 disease (61.8%) and N1 nodal involvement (58.8%) than did the AC group (T4 disease: 34.0%, P = 0.001; N1 involvement: 39.0%, P = 0.02). Patients in the ASC-SCC group who underwent R0 resections had significantly better 1-year survival (30%) than those who underwent R1 or R2 resections (0%; P = 0.025), but lower 1-year survival rates than similar-staged patients in the AC group (69.3%; P = 0.016). CONCLUSIONS: Patients with gallbladder ASC-SCC were similar to those with AC in clinical characteristics, but tended to have more infiltration of multiple adjacent organs and lymphatic metastasis. Curative resection could give these patients better outcomes.

Concomitant Precise Hemihepatectomy to Improve the Efficacy of Surgical Treatment for Hilar Cholangiocarcinoma: Analysis of 38 Cases.

UNLABELLED: Background/Aims: Hilar cholangiocarcinoma (HC) is associated with low rates of resectability and curability, high morbidity and mortality, and poor long-term survival. Radical tumor resection with negative surgical margins provides the only chance of cure and long-term survival. The present study was to investigate the efficacy of concomitant precise hemihepatectomy for HC. METHODOLOGY: The clinical data of 38 patients who underwent surgery for HC with concomitant precise hemihepatectomy at our center from January 2009 to October 2012 were analyzed retrospectively. Survival curves were generated using the Kaplan-Meier method. Univariate and multivariate analysis were performed using the Cox proportional hazards model. RESULTS: R0 resection was performed in 32 patients (84.2%), R1 resection in 4 (10.5%), and R2 resection in 2 (5.3%). Two patients died during the perioperative period (mortality rate 5.3%). The most common postoperative complications were bile leakage (28.9%, 11/38) and hepatic dysfunction (21.1%, 8/38). The overall 1-, 2-, and 3-year survival rates were 65.8%, 36.8%, and 21.1%, respectively. The median survival time was 22.0 months. There were significant differences in survival between R0 and R1/R2 resection (χ2 = 4.516, P < 0.05) and between N0 and N1/N2 disease (χ2 = 10.397, P < 0.05). Univariate and multivariate analysis identified a positive surgical margin, lymph node metastasis and hepatic artery resection as prognostic indicators. CONCLUSIONS: Concomitant precise hemihepatectomy significantly improves the efficacy of radical surgical resection for HC. Precise liver resection, preservation of the hepatic artery, and selective preoperative biliary drainage are important to minimize postoperative morbidity and mortality.

Healing of stoma after magnetic biliary-enteric anastomosis in canine peritonitis models.

OBJECTIVE: To assese the healing of stoma after magnetic anastomosis for the reconstruction of biliary-enteric continuity under severe inflammation. METHODS: Acute bile duct injury was constructed as a bile peritonitis model in mongrel dogs (n=32). Magnetic anastomosis (group A, n=16) and traditional suture anastomosis (group B, n=16) were performed to reconstruct the biliary-enteric continuity in one stage. Half of the dogs in each group were euthanized on the 30th postoperative day, and the other half on the 90th postoperative day to harvest the stoma region. The healing conditions of the stoma after the 2 anastomotic approaches were observed with naked eyes, under light microscope and scanning electron microscope. RESULTS: The stoma leakage rate (50% versus 0% on the 30th postoperative day, 37.5% versus 12.5% on the 90th postoperative day, both P<0.05) and stenosis degree (13.9%±0.3% versus 7.1%±0.3% on the 30th postoperative day, 17.2%±0.4% versus 9.4%±0.4% on the 90th postoperative day, both P<0.01) were significantly higher in group B than in group A. Compared with traditional manual anastomoses, the histological analysis under light and electron microscope showed a more continuous stoma with more regular epithelium proliferation and collagen arrangement, less inflammation in group A. CONCLUSIONS: Magnetic anastomosis stent ensures better healing of the stoma even under the circumstance of severe inflammation.

Clinical feasibility of laparoscopic left lateral segment liver resection with magnetic anchor technique: The first clinical study from China.

BACKGROUND: Magnetic anchor technique (MAT) has been applied in laparoscopic cholecystectomy and laparoscopic appendectomy, but has not been reported in laparoscopic partial hepatectomy. AIM: To evaluate the feasibility of the MAT in laparoscopic left lateral segment liver resection. METHODS: Retrospective analysis was conducted on the clinical data of eight patients who underwent laparoscopic left lateral segment liver resection assisted by MAT in our department from July 2020 to November 2021. The Y-Z magnetic anchor devices (Y-Z MADs) was independently designed and developed by the author of this paper, which consists of the anchor magnet and magnetic grasping apparatus. Surgical time, intraoperative blood loss, intraoperative accidents, operator experience, postoperative incision pain score, postoperative complications, and other indicators were evaluated and analyzed. RESULTS: All eight patients underwent a MAT-assisted laparoscopic left lateral segment liver resection, including three patients undertaking conventional 5-port and five patients having a transumbilical single-port operation. The mean operation time was 138 ± 34.32 min (range 95-185 min) and the mean intraoperative blood loss was 123 ± 88.60 mL (range 20-300 mL). No adverse events occurred during the operation. The Y-Z MADs showed good workability and maneuverability in both tissue and organ exposure. In particular, the operators did not experience either a "chopstick" or "sword-fight" effect in the single-port laparoscopic operation. CONCLUSION: The results show that the MAT is safe and feasible for laparoscopic left lateral segment liver resection, especially, exhibits its unique abettance for transumbilical single-port laparoscopic left lateral segment liver resection.

Who benefits from R0 resection? A single-center analysis of patients with stage â £ gallbladder cancer.

OBJECTIVES: Most patients with gallbladder cancer (GBC) present with advanced-stage disease and have a poor prognosis. Radical resection remains the only therapeutic option to improve survival in patients with GBC. This study aimed to analyze the prognostic factors in patients with stage Ⅳ GBC and to identify a subgroup of patients who might benefit from R0 resection. METHODS: A total of 285 patients with stage Ⅳ GBC were retrospectively analyzed at our institution from January 2008 to December 2012. Factors potentially influencing the prognosis of GBC after surgery were analyzed by univariate and multivariate analyses. RESULTS: The 1-, 3-, and 5-year overall survival rates were 6.6% (15/229), 0.9% (2/229), and 0 (0/229), respectively. Ascites (relative risk [RR] = 1.631, 95% confidence interval [CI]: 1.221-2.180, P = 0.001), pathological grade (RR = 1.337, 95% CI: 1.050-1.702, P = 0.018), T stage (RR = 1.421, 95% CI: 1.099-1.837, P = 0.000), M stage (RR = 1.896, 95% CI: 1.409-2.552, P = 0.000), and surgery (RR = 1.542, 95% CI: 1.022-2.327, P = 0.039) were identified as independent risk factors influencing prognosis. The median survival time (MST) was significantly higher in patients undergoing R0 resection than in those undergoing R1/R2 resection (6.0 vs. 2.7 months; P < 0.001). In subgroup analyses, stage ⅣA patients benefited from R0 resection (MST for R0 vs. R1/R2, 11.0 vs. 4.0 months; P = 0.003), while R0 resection had a significant survival benefit than R1/R2 resection in patient with stage ⅣB GBC without distant metastasis (MST for R0 vs. R1/R2, 6.0 vs. 3.0 months; P = 0.007). CONCLUSION: Ascites, pathological grade, T stage, M stage, and surgery were independent risk factors influencing prognosis in patients with stage IV GBC. N2 lymph node metastasis did not preclude curative resection, and radical resection should be considered in patients with stage Ⅳ GBC without distant metastasis once R0 margin was achieved.

Estimating survival benefit of adjuvant therapy based on a Bayesian network prediction model in curatively resected advanced gallbladder adenocarcinoma.

BACKGROUND: The factors affecting the prognosis and role of adjuvant therapy in advanced gallbladder carcinoma (GBC) after curative resection remain unclear. AIM: To provide a survival prediction model to patients with GBC as well as to identify the role of adjuvant therapy. METHODS: Patients with curatively resected advanced gallbladder adenocarcinoma (T3 and T4) were selected from the Surveillance, Epidemiology, and End Results database between 2004 and 2015. A survival prediction model based on Bayesian network (BN) was constructed using the tree-augmented naïve Bayes algorithm, and composite importance measures were applied to rank the influence of factors on survival. The dataset was divided into a training dataset to establish the BN model and a testing dataset to test the model randomly at a ratio of 7:3. The confusion matrix and receiver operating characteristic curve were used to evaluate the model accuracy. RESULTS: A total of 818 patients met the inclusion criteria. The median survival time was 9.0 mo. The accuracy of BN model was 69.67%, and the area under the curve value for the testing dataset was 77.72%. Adjuvant radiation, adjuvant chemotherapy (CTx), T stage, scope of regional lymph node surgery, and radiation sequence were ranked as the top five prognostic factors. A survival prediction table was established based on T stage, N stage, adjuvant radiotherapy (XRT), and CTx. The distribution of the survival time (>9.0 mo) was affected by different treatments with the order of adjuvant chemoradiotherapy (cXRT) > adjuvant radiation > adjuvant chemotherapy > surgery alone. For patients with node-positive disease, the larger benefit predicted by the model is adjuvant chemoradiotherapy. The survival analysis showed that there was a significant difference among the different adjuvant therapy groups (log rank, surgery alone vs CTx, P < 0.001; surgery alone vs XRT, P = 0.014; surgery alone vs cXRT, P < 0.001). CONCLUSION: The BN-based survival prediction model can be used as a decision-making support tool for advanced GBC patients. Adjuvant chemoradiotherapy is expected to improve the survival significantly for patients with node-positive disease.

Role of transforming growth factor-beta signaling pathway in pathogenesis of benign biliary stricture.

AIM: To characterize the expression of members of the transforming growth factor-beta (TGF-beta)/Smad/connective tissue growth factor (CTGF) signaling pathway in the tissue of benign biliary stricture, and to investigate the effect of TGF-beta signaling pathway in the pathogenesis of benign biliary stricture. METHODS: Paraffin embedded materials from 23 cases of benign biliary stricture were analyzed for members of the TGF-beta/Smad/CTGF signaling pathway. TGF-beta (1), TbetaR I , TbetaR II , Smad4, Smad7 and CTGF protein were detected by immunohistochemical strepto-advidinbiotin complex method, and CTGF mRNA was evaluated by hybridization in situ, while 6 cases of normal bile duct served as controls. The percentages of positive cells were counted. The correlation between TGF-beta (1), Smad4 and CTGF was analyzed. RESULTS: The positive expression ratios of TGF-beta (1), TbetaR I , TbetaR II , Smad4, CTGF and CTGF mRNA in 23 cases with benign biliary stricture were 91.3%, 82.6%, 87.0%, 78.3%, 82.6% and 65.2%, respectively, significantly higher than that in 6 cases of normal bile duct respectively (vs 33.3%, 16.7%, 50.0%, 33.3%, 50.0%, 16.7%, respectively, P < 0.05). The positive expression ratio of Smad7 in cases with benign biliary stricture was 70.0%, higher than that in normal bile duct, but this difference is not statistically significant 70.0% vs 50%, P > 0.05). There was a positive correlation between positive expression of TGF-beta (1), Smad4 and CTGF in cases with benign biliary stricture. CONCLUSION: The high expression of TGF-beta/Smad/CTGF signaling pathway plays an important role in the pathogenesis of benign biliary stricture.

Neuropilin-1 promotes primary liver cancer progression by potentiating the activity of hepatic stellate cells.

As a co-receptor for a variety of cytokines, neuropilin-1 (NRP-1) is detectable in primary liver cancer (PLC) cells. Previous studies determined that silencing of NRP-1 expression attenuated the proliferation, migration and invasion of PLC cells. An increasing number of studies have highlighted the crucial role of the tumor microenvironment in the pathogenesis of cancer. Hepatic stellate cells (HSCs) are one of the major interstitial cell types present in the liver tumor microenvironment, and can promote the proliferation, migration and invasion of PLC cells. It remains unknown whether NRP-1 can promote PLC progression by potentiating the activity of HSCs. In the present study, the expression of NRP-1, and its co-expression with platelet-derived growth factor receptor-ß, in HSCs was detected via immunofluorescence. LX2 HSCs were transfected with NRP-1 short hairpin RNA lentiviral vectors and their proliferation was observed. The proliferation, migration and invasion of HepG2 cells co-cultured with LX2 cells were also observed. Finally, LX2 and HepG2 cells were co-injected into nude mice as subcutaneous xenografts, and the tumor growth and α-smooth muscle actin expression levels were observed. NRP-1 knockdown attenuated LX2 cell activation, with concomitant downregulation of HepG2 cell proliferation, migration and invasion (P<0.05). Thus, silencing of NRP-1 expression may inhibit the activation of HSCs, as well as the proliferation, migration and invasion of PLC cells. The mechanism underlying the inhibition of PLC cell progression is possibly mediated by the inhibition of HSC activation, reduction of transforming growth factor-ß1 levels in the conditioned medium and downregulation of extracellular signal-related kinase activity in PLC cells. Thus, NRP-1 could be regarded as a potential gene therapy target for PLC.

Analysis of prognostic factors for survival after surgery for gallbladder cancer based on a Bayesian network.

The factors underlying prognosis for gallbladder cancer (GBC) remain unclear. This study combines the Bayesian network (BN) with importance measures to identify the key factors that influence GBC patient survival time. A dataset of 366 patients who underwent surgical treatment for GBC was employed to establish and test a BN model using BayesiaLab software. A tree-augmented naïve Bayes method was also used to mine relationships between factors. Composite importance measures were applied to rank the influence of factors on survival time. The accuracy of BN model was 81.15%. For patients with long survival time (>6 months), the true-positive rate of the model was 77.78% and the false-positive rate was 15.25%. According to the built BN model, the sex, age, and pathological type were independent factors for survival of GBC patients. The N stage, liver infiltration, T stage, M stage, and surgical type were dependent variables for survival time prediction. Surgical type and TNM stages were identified as the most significant factors for the prognosis of GBC based on the analysis results of importance measures.

Clinical epidemiological survey of gallbladder carcinoma in northwestern China, 2009-2013: 2379 cases in 17 centers.

OBJECTIVE: To analyze the clinical epidemiological characteristics of patients with gallbladder carcinoma recruited from 17 hospitals in five northwestern provinces of China (Shaanxi Province, Gansu Province, Qinghai Province, Ningxia Hui Autonomous Region, and Xinjiang Uygur Autonomous Region) from 2009 to 2013, and to summarize the clinical diagnosis and treatment data of gallbladder carcinoma. METHODS: Clinical information of 2379 patients with gallbladder carcinoma from 17 hospitals in five northwestern provinces of China was retrospectively collected and analyzed using the "Questionnaire for Gallbladder Carcinoma Patients in Northwestern Area of China." All information was verified with EpiData software and analyzed with SPSS 13.0 software. RESULTS: (1) Gallbladder carcinoma accounted for 2.7% (2379/86,609) of all biliary tract diseases during the study period, which was significantly higher than that from 1986 to 1998 (P < 0.001). (2) Gallbladder carcinoma was more prone to occur in elderly women. The male:female incidence ratio was 1.0:2.1, the average age of onset of disease was 63.7 ± 11.3 years, and the incidence was higher in farmers than in other occupational groups. (3) A total of 57.2% (1360/2379) of patients with gallbladder carcinoma also had gallstones. (4) Abdominal pain (1796/2379, 75.5%) and jaundice (727/2379, 30.6%) were the most common clinical manifestations, 81.2% (1527/1881) were positive in those receiving B ultrasound examinations and 90.7% (1567/1727) were positive in those undergoing computed tomography, and 64.5% (1124/1742) of patients with gallbladder carcinoma were positive for carbohydrate antigen (CA) 19-9. (5) The pathological type of gallbladder carcinoma was mainly moderately and poorly differentiated adenocarcinoma with a high degree of malignancy. At admission, 55.1% (1091/1981) of patients had stage IV cancer among patients with TNM staging information; 55.9% (1331/2379) had lymphatic metastasis, 29.7% (706/2379) had bile duct metastasis, and 53.1% (1263/2379) had liver metastasis. (6) A total of 283 patients (283/2379, 11.9%) had incidentally detected gallbladder carcinoma. (7) The rate of radical surgical resection was 30.4% (723/2379). CONCLUSION: The proportion of gallbladder carcinoma in biliary tract diseases in the northwestern area of China was significantly higher from 2009 to 2013 than from 1986 to 1998. Gallbladder carcinoma was common in older women and mainly diagnosed at an advanced stage. Compared with other surveys in different regions, the rate of metastasis in this survey was high, leading to a low resection rate. Populations at high risk should undergo B-ultrasound examinations at regular follow-up intervals to increase the rate of early diagnosis of gallbladder carcinoma.

Mechanism of benign biliary stricture: a morphological and immunohistochemical study.

AIM: To explore the mechanism of benign biliary stricture. METHODS: A model of trauma of bile duct was established in 28 dogs. The anastomosed tissues were resected and examined by light and electron microscopes on day 3, in wk 1, 3 and mo 3, 6 after operation. CD68, TGF-beta1 and alpha-SMA were examined by immunohistochemical staining, respectively. RESULTS: The mucosal epithelium of the bile duct was slowly recovered, chronic inflammation lasted for a long time, fibroblasts proliferated actively, extracellular matrix was over-deposited. Myofibroblasts functioned actively and lasted through the whole process. The expression of macrophages in lamina propria under mucosa, TGF-beta1 in granulation tissue, fibroblasts and endothelial cells of blood vessels, alpha-SMA in myofibroblasts were rather strong from the 1st wk to the 6th mo after operation. CONCLUSION: The type of healing occurring in bile duct belongs to overhealing. Myofibroblasts are the main cause for scar contracture and stricture of bile duct. High expressions of CD68, TGF-beta1 and alpha-SMA are closely related to the active proliferation of fibroblasts, extracellular matrix over-deposition and scar contracture of bile duct.

Analysis of prognostic factors for survival after hepatectomy for hepatocellular carcinoma based on a bayesian network.

BACKGROUND: The prognosis of hepatocellular carcinoma (HCC) after hepatectomy involves many factors. Previous studies have evaluated the separate influences of single factors; few have considered the combined influence of various factors. This paper combines the Bayesian network (BN) with importance measures to identify key factors that have significant effects on survival time. METHODS: A dataset of 299 patients with HCC after hepatectomy was studied to establish a BN using a tree-augmented naïve Bayes algorithm that could mine relationships between factors. The composite importance measure was applied to rank the impact of factors on survival time. RESULTS: 124 patients (>10 months) and 77 patients (≤10 months) were correctly classified. The accuracy of BN model was 67.2%. For patients with long survival time (>10 months), the true-positive rate of the model was 83.22% and the false-positive rate was 48.67%. According to the model, the preoperative alpha fetoprotein (AFP) level and postoperative performance of transcatheter arterial chemoembolization (TACE) were independent factors for survival of HCC patients. The grade of preoperative liver function reflected the tendency for postoperative complications. Intraoperative blood loss, tumor size, portal vein tumor thrombosis (PVTT), time of clamping the porta hepatis, tumor number, operative method, and metastasis were dependent variables in survival time prediction. PVTT was considered the most significant for the prognosis of survival time. CONCLUSIONS: Using the BN and importance measures, PVTT was identified as the most significant predictor of survival time for patients with HCC after hepatectomy.

Relationship between inducible nitric oxide synthase expression and angiogenesis in primary gallbladder carcinoma tissue.

AIM: To explore the relationship between angiogenesis and biological behaviors of primary gallbladder carcinoma (PGBC), the relationship between the expression of inducible nitric oxide synthase (iNOS) and biological behaviors of PGBC and its relationship with the expression of iNOS and angiogenesis of PGBC. METHODS: The expression of iNOS and micro-vessel density (MVD) were assessed by immunohistochemical method and image analysis system in 40 specimens of PGBC and in 8 specimens of normal gallbladder. The immunostaining results and related clinicopathologic materials were analyzed by statistical methods. RESULTS: MVD in PGBC was significantly higher than that in normal gallbladder tissue (46+/-14 vs 14+/-6, P<0.05), and was not related with age, gender, tumor size and histological type. MVD of poorly and undifferentiated tumor tissues was higher than that of moderately-differentiated and well-differentiated tumor tissues (52+/-9 vs 43+/-9 vs 33+/-6, P<0.01). MVD of Nevin IV and V stages was higher than that of Nevin I, II and III stages (52+/-8 vs 37+/-13, P<0.01). MVD of cases with lymphatic or liver metastasis was significantly higher than that without liver metastasis (55+/-6 vs 42+/-10, P<0.05)or lymphatic metastasis (53+/-8 vs 38+/-8, P<0.01). The positive level index (PLI) of iNOS in PGBC was 0.435+/-0.134, and was not related with age, gender, tumor size, histological type, differentiation and clinical stage of PGBC. The PLI of iNOS in cases with lymphatic metastasis was higher than that without lymphatic metastasis (0.573+/-0.078 vs 0.367+/-0.064, P<0.01). The PLI of iNOS in cases with liver metastasis was higher than that without liver metastasis (0.533+/-0.067 vs 0.424+/-0.084, P<0.05). There was a significant correlation between PLI of iNOS and MVD in PGBC (P<0.05). CONCLUSION: Angiogenesis of PGBC is significantly related to the biological behaviors of PGBC. The expression of iNOS is related to the biological behaviors of PGBC. The detection of MVD and the expression of iNOS in PGBC can be used as parameters to determine the degree of malignancy and prognosis.

Microstructural and ultrastructural changes in the healing process of bile duct trauma.

OBJECTIVE: To explore the mechanism of benign biliary stricture caused by bile duct trauma. METHODS: A model of trauma of the common bile duct was established in 28 dogs and then repaired. The anastomotic tissues were taken on 3 days, 1 week, 3 weeks, 3 months, and 6 months respectively after operation and examined by using light microscopy and electromicroscopy. Macrophage, transforming growth factor beta 1 (TGF-beta1) and alpha-smooth muscle actin (alpha-SMA) were studied immunohistochemically. RESULTS: The mucosal epithelium of the common bile duct restored poorly, chronic inflammation lasted for a long time, fibroblasts proliferated actively, extracellular matrix overdeposited, and myofibroblasts functioned actively during the whole healing process. Immunohistochemical test showed a high expression of macrophage, TGF-beta1 and alpha-SMA during the healing process lasting a long duration. Macrophages were found in the lamina propria under mucosa, TGF-beta1 in the granular tissue, fibroblasts and endothelial cells of blood vessels, while alpha-SMA in the myofibroblasts and smooth muscle tissue. CONCLUSIONS: The healing of the bile duct is in the mode of overhealing. Myofibroblast is the main cause for contracture of scar and stricture of the bile duct. The high expression of macrophage, TGF-beta1 and alpha-SMA is closely related to active proliferation of fibroblasts, extracellular matrix overdeposition and scar contracture of the bile duct.

Reoperation for benign biliary tract diseases in 149 cases: causes and prevention.

BACKGROUND: Failure to diagnose and treat benign biliary tract disease relatively common surgical disease may cause serious consequences. Since the introduction of B-mode ultrasonography, CT, or MRI early and accurate diagnosis of the disease has been possible. In clinical practice, however, these methods have not been adequately used. Inappropriate surgical procedures can also lead to bile duct injury or stenosis after injury, residual cholecystitis, stenosis after cholangiojejunostomy, or stenosis of the Oddi's sphincter. But improvement of the diagnosis and treatment of benign biliary tract disease remains a great challenge to clinicians. METHODS: A total of 149 patients with benign biliary tract disease who had received reoperation from June 1988 to June 2001 were analyzed retrospectively. Among them 95 patients (63.76%) received operation twice and 38 (25.5%) underwent 3 operations. Sixteen patients (10.74%) needed 4 or more operations. The procedures for the first operation included cholecystectomy (71 patients, 47.65%), cholecystectomy with exploration of the common bile duct (42, 28.19%), cholangiojejunostomy (21, 14.1%), and laparoscopic cholecystectomy (15, 10.06%). RESULTS: The causes for reoperation included residual and recurrent bile duct stones in 53 patients (35.57%), bile duct injury or stenosis after injury in 41 (27.52%), residual cholecystitis with or without stones in 28 (18.8%), stenosis after cholangiojejunostomy in 17 (11.41%), stenosis of the Oddi's sphincter in 5 (5.35%), and others in 5 (5.35%). Four patients (2.68%) died after operation. CONCLUSIONS: To prevent reoperation for benign biliary tract diseases, the following measures should be taken to increase preoperative diagnostic rate, to understand conditions of the biliary tract by using imaging techniques and cholangiography, to examine comprehensively and carefully with choledochoscopy, cholangiography and B-mode ultrasonography intraoperatively, to choose appropriate operative procedures to decrease the rate of residual stones, and to decide the time for the first repair according to injury type of the bile duct. Roux-en-Y hepaticojejunostomy with cholangioplasty is the best operation for the reconstruction of the biliary tract.

Activated human hepatic stellate cells promote growth of human hepatocellular carcinoma in a subcutaneous xenograft nude mouse model.

Tumor cell microenvironment defines cancer development, also in hepatocellular carcinoma (HCC). Hepatic stellate cells (HSCs) are believed to be the key contributors to tumor microenvironment in HCC, yet their precise role in cancer progression is still unclear. The aim of this study was to determine the effect of human HSCs on progression of HCC using a subcutaneous xenograft nude mouse model. Nude mice were stratified to receive subcutaneous injections of human HCC cell line HepG2 and human HSC line LX-2 (HepG2 + LX-2), HepG2 alone, LX-2 alone, or phosphate-buffered saline. Tumor growth was assessed by measuring tumor size. After 30 days, final tumor size, weight, and histology were assessed. Compared with mice that were only injected HepG2 cells, mice injected with HepG2 + LX-2 exhibited more rapid tumor growth, increased tumor size and weight, higher tumor cell numbers due to increased proliferation and reduced apoptosis, increased fibrotic bands containing LX-2 cells, and increased tumor angiogenesis. In conclusion, HSCs play a significant role in promotion of HCC growth.