Colorectal cancer derived organotypic spheroids maintain essential tissue characteristics but adapt their metabolism in culture.

BACKGROUND: Organotypic tumor spheroids, a 3D in vitro model derived from patient tumor material, preserve tissue heterogeneity and retain structural tissue elements, thus replicating the in vivo tumor more closely than commonly used 2D and 3D cell line models. Such structures harbour tumorigenic cells, as revealed by xenograft implantation studies in animal models and maintain the genetic makeup of the original tumor material. The aim of our work was a morphological and proteomic characterization of organotypic spheroids derived from colorectal cancer tissue in order to get insight into their composition and associated biology. RESULTS: Morphological analysis showed that spheroids were of about 250 µm in size and varied in structure, while the spheroid cells differed in shape and size and were tightly packed together by desmosomes and tight junctions. Our proteomic data revealed significant alterations in protein expression in organotypic tumor spheroids cultured as primary explants compared to primary colorectal cancer tissue. Components underlying cellular and tissue architecture were changed; nuclear DNA/ chromatin maintenance systems were up-regulated, whereas various mitochondrial components were down-regulated in spheroids. Most interestingly, the mesenchymal cells appear to be substantial component in such cellular assemblies. Thus the observed changes may partly occur in this cellular compartment. Finally, in the proteomics analysis stem cell-like characteristics were observed within the spheroid cellular assembly, reflected by accumulation of Alcam, Ctnnb1, Aldh1, Gpx2, and CD166. These findings were underlined by IHC analysis of Ctnnb1, CD24 and CD44, therefore warranting closer investigation of the tumorigenic compartment in this 3D culture model for tumor tissue. CONCLUSIONS: Our analysis of organotypic CRC tumor spheroids has identified biological processes associated with a mixture of cell types and states, including protein markers for mesenchymal and stem-like cells. This 3D tumor model in which tumor heterogeneity is preserved may represent an advantageous model system to investigate novel therapeutic approaches.

Pre-operative anaemia in colon cancer patients became normal after more than a year post-operatively but did not influence oncological outcome in the final analysis.

BACKGROUND: Pre-operative anaemia has been related to adverse outcomes after surgical management of colorectal cancer. How various factors may contribute to anaemia and also its post-operative recovery has not been extensively investigated. METHODS: Two hundred and thirty five colon cancer patients treated surgically in a community teaching hospital in Norway between 2007 and 2009 were prospectively examined. RESULTS: Anaemia was detected in 53.8%, which was dependent on tumour location, albumin level and female gender. More than 60% of all patients were overweight or obese. Pre-operative BMI was negatively associated with age and positively associated with albumin level (p = 0.018 for both). Haemoglobin levels slowly returned to normal during the first year in significant incremental steps, coincidental with an increase in BMI. At 6 months post-operative, age, albumin, female gender and TNM stage (p = 0.010; p = 0.020; p < 0.001; p = 0.028, respectively) were significantly connected with anaemia, whereas only age and albumin (p = 0.016; p = 0.004, respectively) were associated with a reduced BMI. Pre-operative anaemia gave a significantly worse overall survival (p = 0.040) in the univariate analysis but was not significant in the multivariate analysis. CONCLUSION: Beside the influence of tumour location, it appeared that pre-operative Hgb had some relation to a reduced nutritional state. The post-operative recovery of anaemia was more closely related to an improved nutritional state. The relation between anaemia and oncological outcome should undergo further studies.

Outcome after introduction of complete mesocolic excision for colon cancer is similar for open and laparoscopic surgical treatments.

BACKGROUND: Complete mesocolic excision (CME) and a high (apical) vascular tie may improve oncologic outcome after surgery for colon cancer. Our primary aim was to emulate a previous national result of 73.8% overall survival (OS) with both the open and laparoscopic techniques. METHODS: A prospective study of radical colon cancer was initiated in a Norwegian community teaching hospital in 2007 and comprised a consecutive group of 251 patients with TNM stages I-III that had surgery according to the CME principle. Oncological outcome was assessed as OS, disease-free survival (DFS) and cancer-specific survival (CSS), as well as time to recurrence (TTR), using Cox regression analysis. RESULTS: In-hospital mortality was 3.6%, 2.3% for laparoscopic surgery and 4.9% for open management. Significantly more patients in the open CME group developed complications in the short term (p < 0.001). Twelve or more lymph nodes were retrieved from 82.9% (208/251) of the specimens. Overall 3-year OS was 84.5%, DFS 77.4%, CSS 91.5% and TTR 86.8%. The surgical approach was not a significant predictor for any of the survival parameters. CONCLUSIONS: There was no survival difference between open and laparoscopic CME colonic resections, and the present OS improved from a previous OS from 2000.

Diagnosis and management of gallbladder calculus disease.

BACKGROUND: The number and rate of cholecystectomy are increasing worldwide, although indications for operative treatment remain empirical, and several issues in the understanding of the condition are not concisely outlined. Our intention is to summarize and interpret current opinion regarding the indications and timing of cholecystectomy in calculous gallbladder disease. METHODS: Publications concerned with gallstone disease and related topics were searched for in MEDLINE using PubMed and summarized according to clinical scenarios with an emphasis on recent research. RESULTS: Only one randomized controlled trial has investigated the management (conservative vs. surgery) of patients with acute cholecystitis and several have compared early with deferred surgery. Two RCTs have examined treatment of uncomplicated, symptomatic gallstone disease. Apart from these, the overwhelming majority of publications are retrospective case series. CONCLUSIONS: Recent literature confirms that cholecystectomy for an asymptomatic or incidental gallstone is not justified. Symptomatic, uncomplicated gallstone disease may be classified into four severity groups based on severity and frequency of pain attacks, which may guide indication for cholecystectomy. Most patients below the age of 70 seem to prefer operative treatment. Acute cholecystitis may be treated with early operation if reduction of hospital days is an issue. Patients older than 70 years with significant comorbidities may forego surgical treatment without undue hazard. Symptoms following cholecystectomy remain in 25% or more and recent evidence suggest these are caused by a functional gastrointestinal disorder.

Increased cancer incidence in some gallstone diseases, and equivocal effect of cholecystectomy: a long-term analysis of cancer and mortality.

BACKGROUND: Our aim was to investigate cancer incidence and the cause of long-term mortality in different gallstone diseases and conditions. STUDY DESIGN: The study population consisted of 2034 subjects: 224 persons diagnosed with asymptomatic gallstones in 1983, 254 patients who underwent cholecystectomy in 1983, and 513 patients with symptomatic uncomplicated gallstones (SGS, n = 337) or acute cholecystitis (AC, n = 176) between 1992 and 1994. One thousand and forty-three people who participated in a population study in 1983 were controls. RESULTS: An overall increased risk of cancer, as well as higher mortality, was found among persons with asymptomatic gallstones compared to controls (HR 1.46, 95% CI: 1.06-2.00 and HR 1.39, 95% CI: 1.08-1.78), whereas patients who underwent cholecystectomy in 1983 showed a slightly higher risk (not significant) for both cancer and death than controls. Among patients with SGS from 1992 to 1994 there was a significantly higher risk of contracting cancer in patients who had undergone surgery (HR = 2.56, 95% CI: 1.13-5.83). For patients with AC, there was no significant difference between surgically treated and non-surgically treated subjects, but there was a higher risk of cancer in all AC compared to SGS patients (HR 2.03, 95% CI: 1.20-3.43). Mortality did not differ significantly between surgically treated and non-surgically treated patients with SGS or AC. CONCLUSION: Gallstone patients had a greater risk than the general population for developing cancer, but this was dependent on the type of gallstone condition and treatment. The effect of cholecystectomy seemed dubious.

Long-term follow-up of a randomized controlled trial of observation versus surgery for acute cholecystitis: non-operative management is an option in some patients.

BACKGROUND: Cholecystectomy is routinely recommended to prevent recurrent disease after an initial episode of acute cholecystitis. Therefore, randomized controlled trials have mainly focused on the timing of surgery, but many patients scheduled for cholecystectomy have deferred surgery with long periods of symptom-free intervals. Our present aim is to examine the long-term feasibility and safety of observation compared with surgery. METHODS: Trial of 64 patients with acute cholecystitis previously randomized to observation or cholecystectomy, which examined outcome in terms of completed randomized treatment and appearance of further symptoms and the need for surgical treatment. Thirty-three patients were randomized to observation and 31 patients to cholecystectomy. Median follow-up was 14 years. RESULTS: Of the 33 patients randomized to observation, 11 (33%) experienced a new event of gallstone-related disease (eight (24.2%) had acute cholecystitis) and 11 (33%) were operated. No significant difference (p = 0.565) was found between the two randomized groups with regard to recurrent disease or complications. Virtually no surgery took place after 5 years of follow-up. The difference in completed randomized treatment between the groups was not significant (p = 0.077). Long-term mortality was equal in those operated and in those observed. CONCLUSIONS: Twenty-four percent of the patients experienced recurrent cholecystitis, but escalation of disease severity or increased mortality was not observed. Long-term observation after acute cholecystitis was feasible in two-thirds of the patients as the risk for recurrent disease was negligible after 5 years.

A 24-year controlled follow-up of patients with silent gallstones showed no long-term risk of symptoms or adverse events leading to cholecystectomy.

BACKGROUND: The fate of asymptomatic gallstones has not been investigated in many studies with a long-term follow-up. We wanted to examine the subsequent rate of cholecystectomy and gallstone-related symptoms in a population examined in 1983. METHODS: Among the persons examined in 1983, unknown (perceived as silent) gallstones were discovered in 20.1% (285/1417) persons. Owing to technical reasons, only 89.9% (1274 persons) of the original study population was retrieved for the present study. Of these, 19.2% (245 persons) had gallstones in 1983 [135 women (55.1%) and 110 men (44.9%), mean age in 1983: 49.9 and 51.3 years, respectively]. RESULTS: Of the 154 still living persons with gallstones from 1983, 134 were traced for follow-up. Of these, 89 underwent a clinical examination and 45 answered a mail or telephone questionnaire. Ultrasonography revealed gallstones in 28.1% (25/89) and 6.7% (9/134) had had the gallbladder removed. 5.5% (5/91) of the deceased patients had had a cholecystectomy. Overall cholecystectomy rate was 6.2%. No link could be shown between the number and the size of gallstones in 1983 and the ultrasonographic demonstration of gallstones in 2007. 43.8% had abdominal pain, and 29.2% had functional abdominal complaints. CONCLUSIONS: Unexpectedly, only a minority of persons examined with ultrasonography had present day gallstones without any obvious explanation for this low figure. The rate of cholecystectomy was low in a conservative setting and no adverse events could be ascertained from such a policy.

Functional abdominal complaints occurred frequently in living liver donors after donation.

BACKGROUND: Donor outcome after living donor liver transplantation has not been examined extensively with regard to postoperative abdominal complaints. We wanted to examine the extent and type of abdominal complaints after removal of a part of the liver and gallbladder in living donors as well as potential similarities with known disorders. METHODS: Twelve patients of mixed ethnicity, nine men, aged 18-45 years, and three women, aged 32-46 years, were enrolled in the study during a 3-year period and followed up at 6 and 12 months. Patients filled out questionnaires pertaining to functional abdominal complaints (FAC) using a recognized questionnaire, Rome II, as well as specific abdominal pain symptoms known from gallstone disease. RESULTS: FAC occurred in 11 patients at 6 months and nine patients at 12 months while abdominal pain occurred in seven and six patients, respectively. Three patients had FAC but no abdominal pain while two patients had no complaints at 12 months. Irritable bowel syndrome (IBS) was found in the majority of patients. CONCLUSIONS: FAC and pain seemed to indicate a general postoperative disorder, of a psychosomatic character, and not connected with removal of part of the liver and gallbladder in particular. However, the occurrence of IBS and FD should merit attention, as they are known to impair quality of life.

Overall survival after resection for colon cancer in a national cohort study was adversely affected by TNM stage, lymph node ratio, gender, and old age.

BACKGROUND: A national surveillance program of colon cancer treatment was introduced in 2007. We examined prognostic factors for colon cancer operated in 2000 with an aim of improving survival in the new program and a special focus on the merit of lymph node yield. METHODS: A cohort of 269 patients, 152 women (56.5%), with a mean age of 71 years, was operated for colon cancer in 2000 at three teaching hospitals and followed up for 7 years. RESULTS: Overall 5-year survival was 58.0%, and overall hospital mortality was 5.2%, with 4.5% in elective cases and 12.5% after urgent surgery. In only 41.1% of the specimens were 12 or more lymph nodes retrieved, but this did not affect survival in the combined cohort, although one of the hospitals achieved a significantly better result with a harvest of 12 or more lymph nodes. In a multivariate analysis, old age, gender, a high lymph node ratio (LNR) at stage III, and tumor-node-metastasis stage were adverse factors for survival. CONCLUSIONS: The operative mortality was high and should be reassessed. The lymph node count did not have a significant impact on outcome overall, whereas the LNR proved significant for stage III. A prospective protocol using overall lymph node yield as a surrogate measure for more radical surgery, nevertheless, seems warranted to improve the lymph node harvest according to international recommendations.

Improved lymph node harvest from resected colon cancer specimens did not cause upstaging from TNM stage II to III.

BACKGROUND: The number of lymph nodes retrieved and examined from a resected colon cancer specimen may be crucial for correct staging. We examined if efforts to increase the lymph node harvest to more than 12 lymph nodes per specimen would upstage some patients from TNM stage II to III. METHODS: Three hospitals compared results from 2000 with those of 2007 in 421 resected patients with stage II and III colon cancer. Hospital A endeavored to improve the surgical procedure while the pathologists enhanced the quality of lymph node sampling. Hospital B did not make any marked changes, while hospital C introduced the GEWF lymph node solvent (glacial acetic acid, ethanol, distilled water, and formaldehyde) in their pathology method. RESULTS: In 2000, 12 or more lymph nodes were harvested in 39.6, 45.0, and 21.1% of the specimens from the three hospitals, while the figures for 2007 were 85.7, 42.0, and 90.3%, respectively. The significant increase in lymph node harvest in two of the hospitals in 2007 compared to 2000 (p < 0.001) did not affect the share of patients with stage III in 2007 (38.7%) compared to 2000 (44.1%) (p = 0.260). The number of positive lymph nodes and the lymph node ratio (LNR) decreased from 2000 to 2007. A lymph node yield of 12 or more was not associated with an increased probability of positive lymph nodes in a multivariable logistic regression analysis. CONCLUSION: More radical surgery and dedicated pathologists and the use of the GEWF solvent significantly increased the lymph node yield but did not upstage patients from TNM stage II to III.

A randomized controlled study of uncomplicated gallstone disease with a 14-year follow-up showed that operation was the preferred treatment.

BACKGROUND/AIMS: Cholecystectomy is considered the treatment of choice for symptomatic gallstone disease. Some patients abstain from surgery and provide the opportunity to study the natural history of cholelithiasis. The aim of the present study was to examine the feasibility and safety of observation after extended long-term follow-up in a randomized controlled trial. METHODS: A total of 137 patients (40.5% of those assessed) were randomized to observation or cholecystectomy and followed up for 14 years. The prevalence of symptomatic events or major complications after treatment was the primary end point. A secondary end point was completion of randomized treatment. RESULTS: There were no differences in outcome between the observation group and the surgical group (p = 0.298). Virtually no cholecystectomy was performed after 5 years of follow-up, and no clear escalation in the severity of the disease was observed. A total of 50.7% of patients from the observation group and 88.2% from the surgical group underwent surgery. The group randomized to surgery completed their designated treatment significantly more often (p < 0.001), especially among patients younger than 70 years of age (p = 0.005). CONCLUSION: Cholecystectomy was the preferred treatment after extended long-term follow-up, but conservative management for symptomatic gallstone disease is an alternative to surgery in the elderly.

Pain persists in many patients five years after removal of the gallbladder: observations from two randomized controlled trials of symptomatic, noncomplicated gallstone disease and acute cholecystitis.

After removal of the gallbladder, pain may persist in some patients. To study this condition, 124 patients from two randomized trials, including those with symptomatic noncomplicated gallbladder stones (n = 90) and acute cholecystitis (n = 34), were interviewed, while 139 patients (90%) excluded from both trials responded to a questionnaire 5 years after the operation. Thirty-four patients (27%) of those randomized had pain; 23 (18%) had diffuse, steady pain; and 11 (9%) had pain attacks resembling their preoperative symptoms. A significant dominance of diffuse pain occurred in women (P = 0.024), especially those younger than 60 years (P = 0.004). A tendency for the diffuse type to be dominant was also present in the group of female patients with symptomatic noncomplicated gallbladder stones (P = 0.052). Of the excluded patients, 18% (25/139) had pain, but 88% of them (96% of the men and 87% of the women) were satisfied with the result of the operation. The overall number of patients with postoperative pain was 22% (59/263). We conclude that persisting abdominal pain 5 years after the operation was mainly of a nonspecific type, found mostly in younger women who had had noncomplicated gallstone disease. Eighty-eight percent of the excluded patients declared themselves satisfied with the result of cholecystectomy.

Successful reconstruction of segmental or subsegmental bile ducts after resection of hilar bile ducts: short- and long-term results.

BACKGROUND/AIMS: Biliodigestive anastomoses to small bile ducts after extensive liver resection or central bile duct remain difficult. The inherent problems associated with this particular procedure have not been widely discussed. In order to demonstrate that reconstruction of segmental or subsegmental bile ducts with mucosa-to-mucosa alignment using transanastomotic stents can reduce morbidity associated with the procedure, we have reviewed our experience with small biliodigestive anastomosis after liver resection. METHODOLOGY: Thirteen patients were operated on from 1995 to 1997. Twelve had malignant diagnoses. A comprehensive standardized technique included the use of short or trans-intestinal trans-anastomotic stents and single layer interrupted absorbable sutures with mucosa-to-mucosa alignment. In the patients, in-hospital morbidity and mortality with focus on bile leaks and long-term signs of cholangitis and stricture formation were investigated with a median observation period of 42 months. RESULTS: Eleven underwent extensive liver resections and two had complicated central resections including the bile duct confluence. The procedures included 40 transected bile ducts and 32 anastomoses to the jejunum. One patient died as a result of rupture of a splenic arterial aneurysm caused by a pancreatic pseudocyst. Other complications which occurred in seven patients were intra-abdominal fluid collection (n=5), wound infection (n=1), minor bile leakage from the liver cut surface (n=1), and pleural effusion (n=4). Postoperatively four patients, who had local recurrence with jaundice, showed cholangitis, whereas the remaining nine patients showed no signs of cholangitis. CONCLUSIONS: Our technique for reconstructing small bile ducts with mucosa-to-mucosa alignment is considered adequate from the viewpoint of short- and long-term results.

The total number of lymph nodes in resected colon cancer specimens is affected by several factors but the lymph node ratio is independent of these.

The number of lymph nodes retrieved from the specimen may be a surrogate measure of the adequacy of extensive colon cancer surgery, but many variables may influence the total lymph node yield of any specimen. We examined which variables would be influential both for negative and positive node sampling.The combined results from 428 patients from three hospitals A to C treated in 2007-2009 with single colon cancers having R0 segmental resections were analysed. The surgical technique and pathology staining methods were slightly different between the hospitals.The mean number of lymph nodes was 15.8 (range 1-60). Twelve or more lymph nodes were harvested in 78% of the specimens. In the multivariate Poisson regression analysis of all TNM stages, the factors associated with the total lymph node harvest were age, pathology handling, tumour location and size (p < 0.001), whereas for TNM stage III alone the pathology handling (p < 0.001) and a radical operating technique (p = 0.003) were highly significant. The total number of lymph nodes was the only significant factor for the number of positive lymph nodes (Posln) according to the multivariate negative regression analysis (p = 0.02) but the analysis of the lymph node ratio (LNR) detected no statistically significant variable.Several factors, and especially the specimen processing technique, were important for the total number of harvested lymph nodes. The number of Posln varied between segments and increased with the total number of harvested lymph nodes, but for LNR no variable was important. LNR seemed to abolish the combined effect of tumour location and the total lymph node yield in prognosis assessment.

Post-cholecystectomy symptoms were caused by persistence of a functional gastrointestinal disorder.

AIM: To classify gallstone disease as a basis for assessment of post-cholecystectomy symptoms. METHODS: One hundred and fifty three patients with a clinical and ultrasonographic diagnosis of gallstones filled out a structured questionnaire on abdominal pain symptoms and functional gastrointestinal disorder (FGID) before and at six months after cholecystectomy. Symptom frequency groups (SFG) were categorized according to frequency of pain attacks. According to certain pain characteristics in gallstone patients, a gallstone symptom score was accorded on a scale from one to ten. A visual analogue scale was used to quantify pain. Operative specimens were examined for size and magnitude of stone contents as well as presence of bacteria. Follow-up took place after six months with either a consultation or via a mailed questionnaire. Results were compared with those obtained pre-operatively to describe and analyze symptomatic outcome. RESULTS: SFG groups were categorized as severe (24.2%), moderate (38.6%), and mild (22.2%) attack frequency, and a chronic pain condition (15%). Pain was cured or improved in about 90% of patients and two-thirds of patients obtained complete symptom relief. Patients with the most frequent pain episodes were less likely to obtain symptom relief. FGID was present in 88% of patients pre-operatively and in 57% post-operatively (P = 0.244). Those that became asymptomatic or improved with regard to pain also had most relief from FGID (P = 0.001). No pre-operative FGID meant almost complete cure. CONCLUSION: Only one third of patients with FGID experienced postoperative relief, indicating that FGID was a dominant cause of post-cholecystectomy symptoms.

Lymph node harvest in colon cancer specimens depends on tumour factors, patients and doctors, but foremost on specimen handling.

There are good indications that the number of lymph nodes found in the specimen after resections for colon cancer somehow has a bearing on prognosis. Many factors have been reported in the literature to influence lymph node retrieval. We wanted to assess these closer with special focus on the pathology handling process in our own practice. A range of international literature was reviewed to study what has been found to influence lymph node harvest. A questionnaire was sent to 13 renowned national and international institutions to explore their handling of the colon cancer specimens to obtain a histological diagnosis. A retrospective, hospital audit was undertaken to examine if the number of lymph nodes and staging after examinations of the specimens varied between individual pathologists. In the literature, tumour and patient characteristics, as well as the surgeon and the pathologist, are found to be influential, but it is difficult to ascertain which ones are truly essential. Fat solvents were found by several to increase the lymph node yield, although some also opposed this finding. Our questionnaire showed some variations in the routines of each Department. A junior pathologist was more likely to inspect the specimen first hand and not more than half employed specific lymph node detection strategies while three of 13 did not seek a minimum number of lymph nodes. Still every department had implemented a standard procedure for such examinations. The internal audit showed without doubt that the devotion of the pathologist secured significantly more lymph nodes from the specimen and this may also have detected more stage III cancers. Several tumour and individual patient characteristics, surgical approach and specimen handling may influence lymph node yield and theoretically, TNM staging. Our investigation specifically suggests that tissue handling by pathologists may be a prominent factor in lymph node harvest from colon cancer specimens.

Increased lymphatic vascular density is seen before colorectal cancers reach stage II and growth factor FGF-2 is downregulated in tumor tissue compared with normal mucosa.

Lymphangiogenesis is an important event in progression of colorectal cancer (CRC), and the estimated lymphatic vascular density (LVD) probably indicates facilitated lymphatic tumor cell invasion and metastasis. However, at what time point during tumor progression this process is triggered, is unclear. The aim of this study was twofold. Firstly, to examine LVD in paired samples of CRC tissue and normal mucosa with specific emphasis on possible difference in LVD between tumors stages II and III, and secondly, the expression of the lymphangiogenic growth factor fibroblast growth factor-2 (FGF-2). Eighteen patients were studied. Immunostaining for podoplanin was performed to highlight lymphatic vessels. FGF-2 mRNA expression was determined by quantitative real-time RT-PCR, whereas protein expression was quantitatively assessed by densitometric analysis of Western blot signal intensity. The immunoblots were further validated by FGF-2 immunostaining of histological sections. LVD was significantly increased in tumor tissue compared with the normal mucosa but no changes in LVD between stages II and III CRC was observed. FGF-2 was found to be downregulated both at the mRNA and protein level in tumor tissues compared with normal mucosa. Lymphangiogenesis was triggered early in tumor development. An increased LVD was established before the tumor reached stage II. FGF-2 was downregulated in tumor tissue. The importance of this finding remains unclear.

Lymphangiogenesis in colorectal cancer--prognostic and therapeutic aspects.

Colorectal cancer (CRC) represents the second most common cause of cancer mortality in the western world. The tumors frequently show metastatic spread which affects different organs such as lymph nodes, liver and lungs. Although the pattern of spread may vary, the initial step usually involves dissemination to regional lymph nodes. At present it is clear that neovessel formation, including lymphangiogenesis, represents key events in tumor progression. However, to what extent lymphangiogenesis contributes in the progression of CRC is unclear. This work focuses on recent progress within the field of tumor lymphangiogenesis with special reference to CRC, and on novel therapeutic strategies for anti-lymphangiogenic therapies. Inhibition of metastatic spread may be achieved by restriction of lymphatic vessel growth by using targeted therapeutic strategies towards molecules involved in lymphangiogenic signalling. Such adjuvant therapeutic approaches in addition to existing therapeutic strategies may represent a favourable treatment for CRCs with higher than average risk of disease recurrence and progression.

Association between gallstone-evoked pain, inflammation and proliferation of nerves in the gallbladder: a possible explanation for clinical differences.

OBJECTIVE: To investigate whether enhanced neuroproliferation could be involved in the pathogenesis of gallstone pain. MATERIAL AND METHODS: Gallbladders from 117 patients with gallstones and 43 controls were examined. The gallbladder samples were immunostained against the pan-neuronal marker PGP 9.5 and the number of nerves and nerve area per tissue area estimated. RESULTS: More nerves and an increased nerve area per tissue area were found in uncomplicated symptomatic gallstone disease. In comparison, acute cholecystitis displayed a significantly (p=0.01) decreased number of nerves and nerve area per tissue area. In both categories, the gallbladder neck contained more nerves (p=0.06 and 0.04, respectively) and an increased nerve area per tissue area (p=0.034 and 0.008, respectively) than the body. CONCLUSIONS: Uncomplicated disease showed enhanced neuroproliferation, significantly more in the gallbladder neck, whereas significantly fewer nerves were observed in acute cholecystitis. Nerve growth alteration may play a role in uncomplicated gallstone pain but the pathology may be different in inflammation.