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.

Clinical significance of P-glycoprotein and glutathione S-transferase π expression in gallbladder carcinoma.

P-glycoprotein (P-gp) and glutathione S-transferase π (GST-π) are not only drug-resistance markers, but also prognostic markers of various cancers. The aim of the present study was to investigate the clinical significance of P-gp and GST-π in gallbladder carcinoma (GBC). Tissue samples from 42 patients with GBC were immunostained. Demographic, clinical and follow-up data were collected and analyzed. The positive expression rates of P-gp and GST-π in the GBC tissues were significantly higher (76.2 and 64.3%, respectively) than that of chronic cholecystitis specimens (30 and 20%, respectively) (P=0.014 and 0.035, respectively), and correlated with the Nevin stage of GBC. Multivariate analysis demonstrated that patients with positive expression of P-gp and GST-π showed a significantly lower 2-year survival rate (11.1 and 12%, respectively) compared with patients with negative expression (55.6 and 45.5%, respectively) (P=0.013 and 0.036, respectively). P-gp was also found to be an independent prognostic marker of 2-year survival rate by logistic regression analysis (B=-2.76, P=0.061). Results of this study suggest that P-gp is a prognostic marker of GBC and the detection of P-gp and GST-π may contribute to the prognosis of GBC and the application of chemotherapy as a therapeutic treatment.

Identification of colitis and cancer in colon biopsies by Fourier Transform Infrared spectroscopy and chemometrics.

Cancer is a disease that does great harms to the health of human beings. FT-IR spectroscopy could identify variability at the molecular level in biological specimens. It is a rapid and noninvasive method, which could be used intraoperatively to modify surgical procedures. The aim of this paper is to identify and separate cancer from colitis in endoscopic colon biopsies through the use of FT-IR spectroscopy. A total of 88 endoscopic colon samples, including 41 cases of colitis and 47 cases of colon cancer, were obtained. Specimens were placed on an ATR accessory linked to FT-IR spectrometer with a MCT detector for greater stability and sensitivity. Later, specimens were sent for the histological examination as the reference in the spectral analysis. 41 colitis and 47 cancer specimens were compared. Spectra preprocessed with smoothing and normalization were used for discrimination analysis. PCA was processed to simplify the spectrum data set. Naive Bayes classifier model was constructed for diagnostic classification. Leave-one-out cross-validation method was utilized to assess the discrimination results. The sensitivity of FT-IR detection for cancer achieves 97.6%. The results showed that colon cancer could be distinguished from colitis with high accuracy using FT-IR spectroscopy and chemometrics.

[Study on estimation of radiation dose using FTIR in mice thymus].

Fourier transform infrared spectroscopy (FTIR) was applied to study the biochemical changes in the radiation damaged mouse thymus which increased with radiation dose and provided a new method for the estimation of the radiation dose of radiation damaged patients. The results demonstrated that with the dose increasing, the peak positions like 1 550, 1 400, 1 400 and 1 640 cm(-1) at the dose of 2, 3 and 5 Gy showed some difference, and there was obvious variance in the intensity: (1) The intensity ratio of 1 085 to 1 236 cm(-1) related to nucleic acid tended to decrease. (2) The intensity ratio of 1 640/1 550 decreased. (3) The intensity at 2 958, 2 925, 1 460 and 1 400 cm(-1) showed no significant difference. The results suggest that it may be possible for FTIR to become an effective method to estimate the radiation dose in clinic.

[FTIR spectroscopic study on gallbladder carcinoma cell and nucleus].

The aim of the present research is to establish the cell basis for the carcinoma tissue diagnosis by exploring a method to obtain the FTIR (Fourier transform infrared spectra) of the cultured carcinoma cell and nucleus with FTIR spectroscopy, and investigating the special spectral features of the carcinoma cell and nucleus compared with the carcinoma tissues. In this paper, the gallbladder carcinoma tissues confirmed by histology were measured using a Nicolet Magna 5700-II FTIR spectrometer and the corresponding FTIR spectra were obtained. The cultured gallbladder carcinoma cell (GBC-SD) and nucleus were centrifuged to provide a small pellet of cell and nucleus for FTIR analysis. The cell and nucleus pellet was then placed on the OMNIC sampler. Then the infrared spectra were recorded by the same equipment. Based on the previously established criteria, a comparative study was subsequently carried out between the spectra of the cultured carcinoma cell and nucleus (GBC-SD) and that of the corresponding gallbladder tissues. Several infrared spectral features of the carcinoma cell and nucleus were obtained. All the results suggest that the spectral features of the carcinoma cell and nucleus can be well reflected by that of the carcinoma tissue, though the later is more complicated, which might originate from the intrinsic complexity of the tissue. This study shows that the diagnosis of carcinoma tissue by FTIR method exhibits sufficient cell basis.

Fourier transform infrared spectroscopy of gallbladder carcinoma cell line.

BACKGROUND: Fourier transform infrared (FT-IR) spectroscopy is a physical method applied to the study of cellular changes at the molecular level in various normal and diseased human tissues, including cancer. This study was undertaken to establish a cellular basis for the diagnosis of carcinoma tissue, using FT-IR spectroscopy to study a carcinoma cell line and investigating the specific spectral features of the cell line. METHODS: The FT-IR spectra of cultured gallbladder carcinoma cells (GBC-SD) smeared on a BaF2 window were measured with a Nicolet Magna750-II FT-IR spectrometer. A comparative study was subsequently carried out between the spectra of cultured gallbladder carcinoma cells and those of corresponding carcinoma tissue. RESULTS: Several infrared spectral features were obtained, and the results suggest that the spectral features of the carcinoma cell line reflect those of carcinoma tissue, though the latter are more complex, probably due to the intrinsic complexity of the tissue. CONCLUSION: The diagnosis of carcinoma tissue by FT-IR spectroscopy has a sufficient cellular basis.

[FTIR spectroscopic study on carcinoma cells].

The aim of this research is to establish the cell basis for the carcinoma tissue diagnosis by exploring a method to obtain the FTIR (Fourier transform infrared) spectra of the cultured carcinoma cells with FTIR spectroscopy and investigating the special spectral features of the carcinoma cells compared with the carcinoma tissues. In the present paper, the gastric carcinoma tissues confirmed by histology were measured using a Nicolet Magna750-II FTIR spectrometer and the corresponding FTIR spectra were obtained. The cultured gastric carcinoma cells (SGC7901) were centrifuged to provide a small pellet of cells for FTIR analysis. The cell pellet was then placed on a specially designed salt plate made of BaF2. Then the infrared spectra were recorded by the same equipment. Based on the previously established criteria, a comparative study was subsequently carried out between the spectra of the cultured carcinoma cells (SGC7901) and that of the corresponding gastric tissues. Several infrared spectral features of the carcinoma cells were obtained: the different bands between cells and tissues locate in the range of 3 000-3 600 cm(-1) and 1 640 cm(-1) which are the range of the hydroxy stretching and blending bands of H2O. There are more H2O out of carcinoma cells in carcinoma tissues, so the strong bands of H2O cover the distinctive bands of carcinoma cells in carcinoma tissues. Although the carcinoma tissue is more complicated, which might originate from the intrinsic complexity of the tissue, the results suggest that the spectral features of the carcinoma cells can be well reflected by that of the carcinoma tissue. This study shows that the diagnosis of carcinoma tissue by FTIR method exhibits sufficient cell basis.

Distribution of natural killer cells and T-lymphocyte subsets in peripheral blood, gallbladder cancer and surrounding tissue.

BACKGROUND: The patient with malignant tumor always show immunologic function drawback and ingravescent with tumor development, especially in the aspect of cell-mediated immunity. This study was undertaken to define the relationship between the immune function of local cells and cancer development by investigating the distribution of natural killer (NK) cells and T-lymphocyte subsets in peripheral blood, the cancer tissue and the tissue surrounding gallbladder carcinoma. METHODS: The numbers of CD4+ and CD8+ T-lymphocytes and NK cells were measured by flow cytometry in samples taken from gallbladder cancer tissue, the surrounding tissues and peripheral blood of 38 patients, and compared with the numbers in the peripheral blood and gallbladder tissue of 30 patients with cholecystitis as controls. RESULTS: The numbers of CD4+ and CD8+ T-cells and NK cells in gallbladder cancer tissues were significantly higher than those in the surrounding tissue and gallbladder with gallstone. However, the ratio of CD4+/CD8+ was lower in the cancer tissue than that in the surrounding tissue and tissue from gallbladders with gallstones. The distribution of CD4+ and CD8+ T-cells and NK cells in mucous membrane of cholecystitis gallbladder and that in the tissue surrounding gallbladder cancer were significantly different. CONCLUSIONS: Disproportionate and imbalanced distribution of NK cells and subsets of T-lymphocytes occurs in the mucous membrane proper of gallbladder cancer and surrounding tissue. Although gallbladder cancer tissue has higher expressions of CD4+, CD8+ and NK cells, the immune function is low or in an inhibited state. In gallbladder cancer immunization therapy, local cellular immunological function should be enhanced and the protective barrier improved.

Analysis of variabilities of serum proteomic spectra in patients with gastric cancer before and after operation.

AIM: To study the variabilities of serum proteomic spectra in patients with gastric cancer before and after operation in order to detect the specific protein markers that can be used for quick diagnosis of gastric cancer. METHODS: Proteomic spectra of 46 serum samples from patients with gastric cancer before and after operation and 40 from normal individuals were generated by IMAC-Cu protein chip and surface-enhanced laser desorption/ ionization time of flight mass spectrometry. RESULTS: Fourteen differentially expressed proteins in serum were screened by analysis of proteomic spectra of preoperative patients and normal individuals. We obtained 4 proteins (heat shock protein 27, glucose-regulated protein, prohibitin, protein disulfide isomerase A3) making up marker pattern which was able to class the patient-team and normal-team. These marker patterns yielded 95.7% sensitivity and 92.5% specificity, respectively. The proteins over-expressed in serum of preoperative patients were obviously down-regulated. CONCLUSION: Specific protein markers of gastric cancer can be used for the quick diagnosis of gastric cancer and judgment of prognosis. SELDI-TOF-MS is a useful tool for the detection and identification of new protein markers in serum.

Use of Fourier-transform infrared spectroscopy to rapidly diagnose gastric endoscopic biopsies.

AIM: To determine if Fourier-transform infrared (FT-IR) spectroscopy of endoscopic biopsies could accurately diagnose gastritis and malignancy. METHODS: A total of 123 gastroscopic samples, including 11 cases of cancerous tissues, 63 cases of chronic atrophic gastritis tissues, 47 cases of chronic superficial gastritis tissues and 2 cases of normal tissues, were obtained from the First Hospital of Xi'an Jiaotong University, China. A modified attenuated total reflectance (ATR) accessory was linked to a WQD-500 FT-IR spectrometer for spectral measurement followed by submission of the samples for pathologic analysis. The spectral characteristics for different types of gastroscopic tissues were summarized and correlated with the corresponding pathologic results. RESULTS: Distinct differences were observed in the FT-IR spectra of normal, atrophic gastritis, superficial gastritis and malignant gastric tissues. The sensitivity of FT-IR for detection of gastric cancer, chronic atrophic gastritis and superficial gastritis was 90.9%, 82.5%, 91.5%, and specificity was 97.3%, 91.7%, 89.5% respectively. CONCLUSION: FT-IR spectroscopy can distinguish gastric inflammation from malignancy.

Association of cyclin D1, p16 and retinoblastoma protein expressions with prognosis and metastasis of gallbladder carcinoma.

AIM: To investigate the role of cyclin D1, p16 and retinoblastoma in cancerous process of gallbladder carcinomas and to assess the relation between cyclin D1, p16, Rb and the biological characteristics of gallbladder carcinoma. METHODS: Forty-one gallbladder carcinoma, 7 gallbladder adenoma and 14 chronic cholecystitis specimens were immunohistochemically and histopathologically investigated for the relation of cyclin D1, p16 and Rb with Nevin staging and pathologic grading. RESULTS: The expression rates of abnormal cyclin D1 in gallbladder carcinoma (68.3%) and gallbladder adenoma (57.1%) were significantly higher than those in chronic cholecystitis (7.1%) (P<0.05). No significant difference was found both among the pathological grades G(1), G(2) and G(3) and among Nevin stagings S(1)-S(2), S(3) and S(4)-S(5) of gallbladder carcinoma. The positive rates of p16 (48.8%) and Rb (58.5%) in gallbladder carcinoma were significantly lower compared to those in adenoma (100.0%) and cholecystitis (100.0%) (P<0.05). The positive rates of p16 and Rb in Nevin stagings S(1)-S(2) (80.0% and 90.0%) and S(3) (46.2% and 61.5%) gallbladder carcinomas were significantly higher than those in S(4)-S(5) (33.3% and 38.8%) (P<0.05), and those in pathologic grades G(1) (54.5% and 81.8%) and G(2) (50.0% and 62.5%) gallbladder carcinoma were significantly higher than those in G(3) (28.6% and 35.7%) (P<0.05). The protein expression of p16 and Rb had a negative-correlation in gallbladder carcinoma (r = -0.2993, P<0.05), and this negative-correlation was correlated with Nevin staging (P<0.05). Moreover, the protein expression of p16 and cyclin D1 had a negative-correlation in gallbladder carcinoma (r = -0.9417, P<0.05). CONCLUSION: Cyclin D1 may play a role in the early stage of gallbladder carcinoma. Mutation of p16 and Rb genes might be correlated with progression of gallbladder carcinoma. Analysis of p16 and Rb can estimate the prognosis of gallbladder carcinoma. Expression of p16 and Rb may be correlated with Nevin staging and pathologic grading in gallbladder carcinoma.

In vivo and in situ detection of colorectal cancer using Fourier transform infrared spectroscopy.

AIM: Real-time and rapid identification of the malignant tissue can be performed during or before surgical operation. Here we aimed to detect in vivo and in situ colorectal cancer by using Fourier transform infrared (FTIR) spectroscopy and fiber-optic technology. METHODS: A total of five patients with large intestine cancer were detected in vivo and in situ. Of them, three cases of colon cancer and one case of cecum cancer were detected intraoperatively and in vivo by using a FTIR spectrometer during surgical operation, and one case of rectum cancer was explored non-invasively and in vivo before the surgical operation. Normal and malignant colorectal tissues were detected in vivo and in situ using FTIR spectroscopy on the basis of fundamental studies. RESULTS: There were significant differences between FTIR spectra of normal and malignant colorectal tissues detected in vivo and in situ. Experimental results revealed that the spectral characteristics of normal and malignant tissues found in vivo and in situ were similar to those obtained from in vitro measurement in our previous fundamental research. CONCLUSION: FTIR fiber-optic attenuated total reflectance (ATR) spectroscopy can identify in situ and in vivo colorectal cancer. FTIR spectroscopic method with fiber optics is a non-invasive, rapid, accurate and in vivo cancer detection technique in clinical diagnosis.

Diagnosis of gastric inflammation and malignancy in endoscopic biopsies based on Fourier transform infrared spectroscopy.

BACKGROUND: Fourier transform infrared (FT-IR) spectroscopy is an effective tool for investigation of chemical changes at the molecular level. We previously demonstrated that FT-IR spectroscopy can reliably distinguish multiple types of carcinoma from healthy tissue. Because various stomach diseases are common, it is important to explore a noninvasive and rapid method to detect malignancy and gastritis in endoscopic biopsies. Our aim was to classify endoscopic biopsies into healthy, gastritis, and malignancy through the use of FT-IR spectroscopy. METHODS: A total of 103 endoscopic samples, including 19 cases of cancer, 35 cases of chronic atrophic gastritis, 29 cases of chronic superficial gastritis, and 20 healthy tissue samples, were obtained at the First Hospital of Xi'an Jiaotong University, China. A modified attenuated total reflectance accessory was linked to a WQD-500 FT-IR spectrometer for biopsy measurement. The spectral characteristics for different types of tissues were correlated with the corresponding pathology results. The gastric biopsies were classified by FT-IR spectroscopy and a discriminant analysis method. RESULTS: There were significant differences in the FT-IR spectra of four types of gastric biopsies. The discriminant analysis results demonstrated that the sensitivity of FT-IR detection for healthy, superficial gastritis, atrophic gastritis, and gastric cancer was 90%, 90%, 66%, 74%, respectively, which could help satisfy clinical diagnostic requirements. CONCLUSION: FT-IR spectroscopy can distinguish disease processes in gastric endoscopic biopsies.

[Novel infrared spectroscopy methods for clinical diagnosis of tumor].

In this paper, the authors have reviewed their investigation on the clinical detection of tumor tissues by infrared spectroscopy in recent ten years. Based on the comparison of different IR spectroscopic methods such as IR transmission spectroscopy, micro-IR spectroscopy etc, the authors found the good consistency of the results of ATR (attenuated total reglection) IR spectroscopic method with those of pathological biopsy. The authors have directly measured the IR spectra of frozen tissues stored in liquid nitrogen and freshly resected tissues, and have realized the measurement of tumor tissues in vito during the operation process using a specially designed IR spectrometer connected with a mid-IR fiberoptic with an ATR probe. The authors have investigated the malignant and normal tissues including parotid, esophagus, stomach, colon, liver, gallbladder, breast, thyroid etc. and compared with the pathological results. The accuracy of this novel IR detection method is more than 90%.

Diagnosis and treatment of polypoid lesions of the gallbladder: report of 194 cases.

BACKGROUND: With the wide use of B-ultrasonography in recent years, the polypoid lesion of the gallbladder (PLG) has been one of the most common diseases detected in biliary surgery. This study was to investigate the diagnostic method and operative indications of PLG. METHODS: The clinical and pathological data of 194 patients with PLG who had received operation at our hospital from January 1994 to September 2002 were analyzed retrospectively. Categorized data were analyzed by the chi-square test. RESULTS: All the patients received preoperative B-ultrasonography. 185 of the 194 PLG patients were diagnosed as having cholecystic polyp, and 9 adenomas. Among the 42 patients who received CT, 6 showed early gallbladder cancer. Pathologically, cholesterol polyps were mostly multiple lesions (64.7%) with a mean diameter of 3.86+/-2.2 mm in 136 patients. Of 16 patients with adenomas, 10 had a tumor diameter of more than 10 mm (62.5%). In 11 patients with gallbladder carcinoma, 7 were accompanied with gallbladder stone (63.6%). In addition, inflammatory polyps and adenomyomas were found in 25 and 6 patients respectively. CONCLUSIONS: B-ultrasonography is the most effective diagnostic method for detecting PLG. When large or irregular lesions are found, CT should be performed in order to avoid missing of gallbladder carcinoma. Operative indications for PLG include: a maximal tumor diameter of more than 10 mm; an over 50-year-old patient with a widebase and a single polyp lesion; a wide-base lesion or a lesion showing a tendency to enlargement; co-existing gallbladder stone or cholecystitis; a patient without other diseases but obvious clinical features and failure of general management; big or long pedicels or polyps at the neck of the gallbladder for preventing the empty of the gallbladder and a history of biliary colic; and PLG with irregularly thickened local gallbladder wall.

Genomic determination of CR1 CD35 density polymorphism on erythrocytes of patients with gallbladder carcinoma.

AIM: To study the changes of quantitative expression, adhering activity and genomic density polymorphism of complement types in erythrocytes (CR1) of patients with gallbladder carcinoma and the related clinical significance. METHODS: Polymerase chain reaction (PCR), Hind III restriction enzyme digestion, quantitative assay of CR1 and adhering activity assay of CR1 in erythrocytes were used. RESULTS: The number and adhering activity of CR1 in patients with gallbladder carcinoma (0.738+/-0.23, 45.9+/-5.7) were significantly lower than those in chronic cholecystitis and cholecystolithiasis (1.078+/-0.21, 55.1+/-5.9) and healthy controls (1.252+/-0.31, 64.2+/-7.4) (P<0.01). The number and adhering activity of CR1 in patients with chronic cholecystitis and cholecystolithiasis (1.078+/-0.21, 55.1+/-5.9) were significantly lower than those in healthy controls (1.252+/-0.31, 64.2+/-7.4) (P<0.05). There was a positive correlation between quantitative expression and adhering activity of CR1 (r = 0.79, P<0.01). Compared with those on preoperative day (0.738+/-0.23, 45.4+/-4.9), the number and adhering activity of CR1 in patients with gallbladder carcinoma decreased greatly on the third postoperative day (0.310+/-0.25, 31.8+/-5.1) (P<0.01), and on the first postoperative week (0.480+/-0.25, 38.9+/-5.2) (P<0.01), but they were increased slightly than those on the preoperative day (P>0.05). The number and adhering activity of CR1 recovered in the second postoperative week(0.740+/-0.24, 46.8+/-5.9) (P<0.01) and increased greatly in the third postoperative week (0.858+/-0.35, 52.7+/-5.8) (P<0.01) in comparison with those on the preoperative day and in the first postoperative week. The number and adhering activity of CR1 of gallbladder carcinoma patients with infiltrating, adjacent lymphogenous and distant organ metastases were significantly lower than those of gallbladder carcinoma patients without them (P<0.01). No difference was observed between the patients with gallbladder carcinoma and healthy individuals in the spot mutation rate of CR1 density gene (chi(2) = 0.521, P>0.05). The distribution of expression was 67.8% in high expression genomic type, 24.8% in moderate expression genomic type, and 7.4% in low expression genomic type. The number and adhering activity of CR1 high expression genomic type gallbladder carcinomas (0.749+/-0.22, 42.1+/-6.2) were significantly lower than those of healthy individuals (1.240+/-0.29, 63.9+/-7.2), and were also significantly lower than those of healthy individuals (0.921+/-0.23, 54.8+/-7.1), but no difference was observed between the number and adhering activity of CR1 lower expression genomic type gallbladder carcinomas (0.582+/-0.18, 44.3+/-5.5) and those of healthy individuals (0.610+/-0.20, 45.8+/-5.7) (P>0.05). CONCLUSION: Defective expression of CR1 in gallbladder carcinoma is mostly acquired through central peripheral mechanisms. The changes in CR1 quantitative expression and adhering activity are consanguineously related to the development and metastasis in gallbladder carcinoma.

Expression of tumor necrosis factor and its receptor in gallstone and gallbladder carcinoma tissue.

BACKGROUND: Some reports have confirmed that occurrence and development of tumor are related with lots of oncogene, anti-oncogene and cytokine. This study was to detect the expression of TNFmRNA, tumor necrosis factor (TNF) and TNF receptor (TNFR) in the gallbladder mucosa which developed from hyperplasia, dysplasia to carcinoma, and to further discuss the pathogenecity of gallbladder carcinoma. METHODS: The expression of TNFmRNA, TNF and TNFR was detected by in situ hybridization and immunohistochemistry on the surgical specimens of hyperplasia, dysplasia and carcinoma of the gallbladder. RESULTS: The percentage of positive cells expressed TNFmRNA in the gallbladder mucosa was 0%, 20%, and 90%, respectively in hyperplasia, dysplasia and carcinoma (P<0.05). The percentage of positive mononuclear cells (MNCs) expressed TNFmRNA in hyperplasia, dysplasia and carcinoma of gallbladder tissues was 15%, 85%, and 90%, respectively (P<0.05). The number of positive MNC high-power field (Hpf) was 4.85+/-1.50, 6.00+/-2.71, and 9.33+/-3.07, respectively (P<0.05). The number of carcinoma cells and MNCs expressed by TNFmRNA per high-power field was increased with the increase of tumor stage. The number of carcinoma cells/Hpf in stages I-III and IV-V was 9.13+/-4.39 and 7.13+/-2.53 (P<0.05), and that of MNC/Hpf was 14.80+/-4.02 and 11.10+/-2.23 (P<0.01). The number of carcinoma cells and MNCs expressed by TNFmRNA per Hpf was increased with the increase of tumor size. In tumors of more than 2 cm or less than 2 cm in diameter, the number of positive carcinoma cells/Hpf was 14.00+/-4.20 and 8.83+/-4.96, respectively (P<0.05), but that of MNC/Hpf was 10.50+/-2.54 and 7.00+/-2.83 (P<0.05). The pattern of TNF protein expression was similar to that of TNFmRNA, whereas TNF protein expression was more frequent and extensive than TNFmRNA expression. TNFR was expressed in endothelial cells and MNCs of carcinoma, and was negative in mucosal epithelial cells and tumor cells. A positive linear correlation in TNFmRNA expression was observed between tumor cells and MNCs (r=0.687, P<0.01), a correlation in TNFmRNA and TNF protein expression of tumor cells (r=0.847, P<0.001), and a correlation in TNFmRNA and TNF protein expression of MNC in tumor tissue (r=0.643, P<0.01). CONCLUSIONS: TNFmRNA and TNF protein expression is increased during the development of gallbladder mucosa from hyperplasia, dysplasia to carcinoma, and is increased with tumor stage. This finding suggests that TNF is involved in the pathogenesis of gallbladder carcinoma induced by gallstones and the TNF expression in cancer cells may serve as a marker for tumor stage.

Utility of serum CA19-9 in diagnosis of cholangiocarcinoma: in comparison with CEA.

AIM: The diagnosis of cholangiocarcinoma is often difficult, making management approaches problematic. A reliable serum marker for cholangiocarcinoma would be a useful diagnostic test. The aims of our study were to evaluate the usefulness of a serum CA19-9 determination in the diagnosis of cholangiocarcinoma. METHODS: We prospectively measured serum CA19-9 and CEA concentrations in patients with cholangiocarcinoma (n=35), benign biliary diseases (n=92), and healthy individuals (n=15). Serum CA19-9 and CEA concentrations were measured by an immunoradiometric assay without knowledge of the clinical diagnosis. RESULTS: The sensitivity of a CA19-9 value >37 KU/L(-1) and a CEA value >22 microg/L(-1) in diagnosing cholangiocarcinoma were 77.14% and 68.57%, respectively. When compared with the benign biliary diseases group, the true negative rates of serum CA19-9 and CEA were 84.78% and 81.52%, respectively. The false positive rates of serum CA19-9 and CEA were 15.22% and 18.48%, whereas the accuracy of serum CA19-9 and CEA were 82.68% and 77.95%, respectively. Serum CA19-9 and CEA concentrations were significantly elevated (P<0.001 and P<0.05) in patients with cholangiocarcinoma (290.31+/-5.34 KU/L(-1) and 36.46+/-18.03 microg/L(-1)) compared with patients with benign biliary diseases (13.38+/-2.59 KU/L(-1) and 13.84+/-3.85 microg/L(-1)) and healthy individuals (12.78+/-3.69 KU/L(-1) and 11.48+/-3.37 microg/L(-1)). In 15 patients undergoing curative resection of cholangiocarcinoma, the mean serum CA19-9 concentration was decreased from a preoperative level of 286.41+/-4.36 KU/L(-1) to a postoperative level of 62.01+/-17.43 KU/L(-1) (P<0.001), and the mean serum CEA concentration from 39.41+/-24.35 microg/L(-1) to 28.69+/-11.03 microg/L(-1) (P<0.05). In patients with cholangiocarcinoma, however, no correlation was found between serum CEA and CA19-9 concentrations (r=0.036). CONCLUSION: These data suggest that the serum CA19-9 determination is a useful addition to the available tests for the differential diagnosis of cholangiocarcinoma. Serum CA19-9 is an effective tumor marker in diagnosing cholangiocarcinoma, deciding whether the tumor has been radically resected and monitoring effect of treatment.

Expression of the bacterial gene in gallbladder carcinoma tissue and bile.

BACKGROUND: The major causive factors of gallbladder carcinoma are very complex. Cholecystitis with gallstone was reported one of the most important factors. Many research revealed that cholecystitis or gallstone can give rise to epithelial hyperplasia of gallbladder mucusa or canceration secondarily. In this study, 46 patients were detected in order to find the relationship between infection of different bacteria and formation of gallbladder carcinoma. METHODS: Using the common gene primer of bacteria 16S ribosomal RNA (rRNA), we detected bacterial gene fragments of gallbladder carcinoma tissues in 46 patients by polymerase chain reaction (PCR). Relative bile was also detected by PCR in 18 patients who underwent operations, including U-tube drainage (1), right or left biliary tube drainage (4), radical cholecystectomy (9), and cholecystorrhaphy (4). The tissue fragments of gallbladder carcinoma from the remaining 28 patients were paraffin slices. RESULTS: The positive rate of bacterial DNA in gallbladder carcinoma tissue was 78.3% (36/46). The sequence of 16S ribosomal RNA gene fragments amplified by PCR was approximately 371 base pairs (bp). Multiple kinds of standard bacterial gene fragments obtained from 36 patients included Colibacillus, B.fragilis, Klebsiella, C.perfringens and Clostridium, with a positive rate of 78.3% (36/46). Among the 36 patients, 14 patients with gallbladder carcinoma received operation and their relative bile at operation was detected bacterial gene fragments with a positive rate of 77.8% (14/18). This result was close to that in gallbladder carcinoma tissues. CONCLUSIONS: Our results suggested that there might be a relationship between occurrence of gallbladder carcinoma and infection of different kinds of bacteria, especially anaerobic bacteria C.perfringens. This reminds us that the gallbladder mucosa stimulated by anaerobic and aerobic bacteria might be the principal cause for the development of carcinoma.