Search Article Keyword  
PubMed Submission Abstract PDF Cited  Click Count: 1364 DownLoad Count: 431 

ISSN 1007-9327 CN 14-1219/R  World J Gastroenterol  2004 December 1;10(23):3480-3484

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

Xing-Yuan Jiao, Ming-De L, Jie-Fu Huang, Li-Jian Liang, Jing-Sen Shi


Xing-Yuan Jiao, Ming-De Lü, Jie-Fu Huang, Li-Jian Liang, Department of Hepatobiliary Surgery, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510080, Guangdong Province, China
Jing-Sen Shi, Hepatobiliary Research Laboratory, First Affiliated Hospital, Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
Supported by National Postdoctor Natural Science Foundation of China, No. 2001-14
Correspondence to: Xing-Yuan Jiao, Department of Hepatobiliary Surgery, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510080, Guangdong Province, China.  jiaoxingyuan@hotmail.com.cn
Telephone: +86-20-34152243    Fax: +86-20-34152456
Received: 2004-01-09    Accepted: 2004-03-16

Abstract
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 (x2 = 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.

Jiao XY, L MD, Huang JF, Liang LJ, Shi JS. Genomic determination of CR1 CD35 density polymorphism on erythrocytes of patients with gallbladder carcinoma. World J Gastroenterol  2004; 10(23): 3480-3484
http://www.wjgnet.com/1007-9327/10/3480.asp


INTRODUCTION
Carcinoma of the gallbladder is the most common neoplasm in biliary tract, and its incidence has been rising in recent years[1-4]. The C3b receptors (CR1, Cd35) on erythrocytes serve as the primary transport system for immune complexes from peripheral blood of the liver[5-8], and CR1 plays an important role in this system, the rate of clearance of immune complexes from the circulation is directly related to the number of CR1 molecules expressed on erythrocytes[9-11]. However, there are few reports on the changes of genomic density polymorphism of complement types in erythrocytes of patients with gallbladder carcinoma. We previously demonstrated the altered levels of cellular immunity and humoral immunity in patients with gallbladder carcinoma, and the effects of radical cholecystectomy on nutritional and immune status in patients with gallbladder carcinoma[12-15]. In this study, we reported the changes in quantitative expression, adhering activity and genomic density polymorphism of complement types in erythrocytes (CR1) of patients with gallbladder carcinoma.

MATERIALS AND METHODS
Patients
A total of 33 patients with gallbladder carcinoma were admitted to the First Affiliated Hospital of Sun Yat-Sen University from August 2000 to August 2002.Those were 16 men and 17 women aged from 50 to 70 years (median, 52.7 years). Nineteen patients were preoperatively diagnosed as polypois, cholecystolithiasis and chronic cholecystitis, and diagnosed as gallbladder carcinoma by frozen sections after surgery. Fourteen patients were diagnosed as gallbladder carcinoma preoperatively, of them, 4 patients had radical cholecystectomy, 7 patients had U-tube drainage operation, 3 patients  cholecystectomy. Direct invasion or distant metastasis was found in 10 patients during operation, simple gallbladder carcinoma in 15 patients, both gallbladder carcinoma and cholecystolithiasis in 18 patients. The patients with gallbladder carcinoma were grouped according to the staging system of Nevin[16], the number of cases on stages I, II, III, IV and V was 7, 11, 5, 7 and 11, respectively. Tumor size ranged from 1.8 cm to 6 cm in diameter (median, 2 cm), 23 patients had tumors smaller than 2 cm in diameter, 10 patients had tumors larger 2 cm in diameter.
      No therapy was administered to patients preoperatively. Meanwhile, 90 patients with cholecystits or cholecystolithiasis had cholecystectomy, including 44 patients with cholecystitis and 46 patients with choleithiasis. No severe systemic diseases were found, such as, myocardial infarction, cerebral vascular accidents, uncontrollable diabetes mellitus, Hypertension. Their disease was stable in a period of one month. All the surgical specimens were examined histologically, and classified into hyperplasia, atypical hyperplasia (mild, moderate and severe) according to Gong et al.[17]. A total of 59 patients exhibited epithelial hyperplasia, 10 patients had mild dysplasia, 10 patients moderate dysplasia, 11 patients severe dysplasia. Normal controls were 30 healthy individuals (18 men and 12 women) aged from 56 years to 72 years (median 61.7 years).

Methods
Quantitative expression, adhering activity and genomic density polymorphism of complement types in erythrocytes (CR1) patients with gallbladder carcinoma were assessed preoperatively (1 wk before surgery), and on d 3, 7, 14 and 21 postoperatively.

Genomic determination of CR1 (Cd35) density polymorphism on erythrocytes[18]
DNA samples  DNA was extracted using the following simplified protocol. Following lysis of erythrocytes, nucleated cells from 10 mL of blood were incubated in 3.5 mL of 10 mmol/L Tris, 2 mmol/L EDTA buffer pH7.5, containing 0.4 mol/L NaCL, 7 g/L sodium dodecylsulfate and proteinase K (30 mg/L) for 16 h at 37 °C, 1 mL of 6 mol/L NaCL was added. After vortexed and ethanol-precipitated, the pellets were washed three times in 700 mL/L ethanol and solubilized in 10 mmol/L Tris, 1 mmol/L EDTA buffer pH7.5.

Sequencing
A construct in pUC-18 containing Hind III-kpn fragments from the introns located within the first and second exons of the short consensus repeat d2 from the long homologous repeat D of CR1 was sequenced. Sequencing was performed using the modified Sanger method with a commercial T7 DNA polymerase kit.

Primers
Primers were designed based on the analysis of internal homologies within the CR1 gene and homologies with other human sequences obtained from the EMBL databank using the Bisance software. The 5’ primer was from base 4 415 to base 4 435 from the CR1 cDNA sequence. The 3’ primer was 75 to 55 from the Hind III monomorphic restriction site in the introns. The latter primer was chosen due to the presence of five mismatches between the CR1 intron sequence and the consensus Hind III 1.9 ku repetitive sequence. Primers were synthesized on a 380 Applied Biosystem apparatus. Twenty base oligonucleotides were recovered from the solid phase by rinsing the column with 250 g/L ammonium. After incubation at 55 °C for 16 h, ammonium was expelled through evaporation using a Speed-Vac apparatus. Primers were used without further purification.

Amplification and agarose gel electrophoresis
PCR was performed using a Techne PHC-I apparatus with the Taq polymerse.
      Amplified DNA was precipitated and then digested using the Hind III restriction enzyme, and separated by electrophoresis in 20 g/L agarose gels. Gels were analyzed under UV illumination. The genomic density polymorphism of complement type 1 (CR1) in erythrocytes was classified into three types: high expression genomic type (1.8 Kb1), moderate expression type (1.8 kb, 1.3 Kb and 1.5 Kb) and low expression genomic type (1.3 Kb, 0.5 Kb).

Quantitative expression of CR1 in erythrocytes of patients with gallbladder carcinoma
EBC enzyme-linked immunosorbent assay  This assay was based on the method of Cornillet et al.[18]. All RBCs in the assay were routinely glutaraldchyde-fixed. Finally, 20 mL of S/BSA were added to each  well, followed by 20 mL of substrate. The plates were incubated at 37 °C for 90 min with gentle agitation every 15-20 min to ensure the maximum contact of RBC with substrate. Subsequently, 100 mL of S/BSA was added to each well, the plate was centrifuged at 1 800 r/min for 90 s and 100 mL of supernatant was transferred to a clean microtitre plate for readings in a plate reader at 405 nm (Dynatech MR5000; DynatchmBillingshurst, Suaaex, UK). In each case, a mean of the readings obtained for the RBC without first antibody (the blank controls) was subtracted from the other readings obtained, in order to correct nonspecific phoshatase-like activity within RBC membranes. Duplicate values were obtained for each RBC sample with each antibody. Black control values were obtained for both the antimouse conjuates and for the anti-rabbit conjuates.

Adhering activity of CR1 in erythrocytes of patients with gallbladder carcinoma
A 50 mL of 1×108/mL RBC suspensions was added to each well, then added and 50 mL of plasma itself and 100 mL of 1×106/mL tumor cells, mixed completely, stained. Fields were examined (×640), 5 RBC combined each tumor cell were labeled as a flower, tumor cell flowers were assessed.

Statistical analysis
For each variable, mupltiple analysis of variance for repeated measurements was used to compare the values measured before operation with those measured at four subsequent time points. The results were presented as mean±SE based on the mixed model of repeated measurement analysis. Statistical analysis was performed with SAS software. P<0.05 was considered statistically significant.

RESULTS
The number and adhering activity of CR1 in patients with gallbladder carcinoma, chronic cholecystitis and cholecystolithiasis and healthy individuals

As shown in Table 1, 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 patients chronic cholecystitis and cholecystolithiasis (1.078±0.21, 55.1±5.9) and healthy individuals controls (1.252±0.31, 64.2±7.4) (P<0.01). In Table 2 the number and adhering activity of CR1 in patients with chronic cholecystitis and cholecystolithiasis were significantly lower than those in healthy controls (P<0.05). There was a positive correlation between quantitative expression and adhering activity of CR1(r = 0.79, P<0.01).
      In the series of epithelial pathologic changes including hyperplasia, atypical hyperplasia, carcinoma in situ or invasive carcinoma, the number and  adhering activity of CR1 decreased gradually (Table 3), and there was a positive correlation between quantitative expression and adhering activity of CR1 (r = 0.77, P<0.01). In Table 4 compared with those on the 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  in 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). In comparison with those on the preoperative day and in the first  postoperative week, 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).
      The number and adhering activity of CR1 in gallbladder carcinoma patients with infiltrating, adjacent lymph node and distant organ metastases were significantly lower than those in gallbladder carcinoma patients without metastasis (Table 5, P<0.01).

Table 1  The number and adhering activity of CR1 in different groups before operation (mean±SE) 

Group Cases CR1 (A405) Adhering activity (%)
Healthy individuals 30 1.252±0.31 64.2±7.4
Chronic cholecystitis 90 1.078±0.21a 55.1±5.9a
Cholecystolithiasis      
Gallbladder carcinoma 33 0.738±0.23bc 45.9±5.7bd

aP<0.05, bP<0.01 vs  normal control; cP<0.05, dP<0.01 vs chronic cholecystitis and cholecystolithiasis.

Table 2  The number and  adhering activity of CR1 in different groups before operation (mean±SE) 

Group Cases CR1(A405) Adhering activity(%)
Healthy individuals 30 1.252±0.31 64.2±7.4
cholecystolithiasis 46 1.027±0.27a 54.4±5.9a
Gallbladder carcinoma with cholecystolithiasis 18 0.731±0.26bc 45.1±6.1bd
Simple gallbladder carcinoma 15 0.728±0.23bc 44.8±6.4bd

aP<0.05, bP<0.01 vs normal control; cP<0.05, dP<0.01 vs chronic cholecystitis.

Table 3
  The number and adhering activity of CR1 in different groups before operation (mean±SE) 

Type Cases CR1(A405) Adhering activity(%)
Simple hyperplasia 59 1.175±0.22 61.1±6.2
Atypical hyperplasia 21 0.906±0.24b 50.8±5.7b
Gallbladder  carcinoma (I-III stage) 23 0.761±0.23b 41.6±6.1b
Gallbladder  carcinoma (IV-V stage) 10 0.602±0.31b 30.4±6.5b
F 6.4 11.5  
P <0.01 <0.01  

bP<0.01 vs each group.

Changes of genomic density polymorphism of CR1 in patients with gallbladder carcinoma
The distribution of genomic density polymorphism of CR1 in patients was 67.8% in high expression genomic type, 24.8% in moderate expression genomic type, 7.4% in low expression genomic type. Compared with healthy controls, no difference was observed between the patients with gallbladder carcinoma and healthy in the spot mutation rate of CR1 density gene (x2 = 0.521, P>0.05).The number and adhering activity of CR1high expression genomic type gallbladder carcinomas (0.749±0.22, 42.1±6.2) were significantly lower than those in healthy individuals (1.240±0.29, 63.9±7.2). The number and adhering activity of CR1 moderate expression genomic type gallbladder carcinomas (0.641±0.19,34.2±5.1) were also significantly lower than those in 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).

Table 4  The number and adhering activity of CR1 on preoperative and postoperative day (mean±SE) 

Group Preoperative 3rd d 1st wk 2nd wk 3rd wk
CR1 (A405) 0.738±0.23 0.310±0.25b 0.480±0.25b 0.740±0.24 0.85±0.32a
Adhering activity (%) 45.4±4.9 31.8±5.1b 38.9±5.2b 46.8±5.9 52.7±5.8a

aP<0.05, bP<0.01 vs normal control.

Table 5
  The number and adhering activity of CR1 in gallbladder carcinoma with adjacent lymph nodes and metastasis (mean±SE)

Group Healthy individu als Infiltrating (-) Infiltrating (+) Adjacent lymph node (-) Adjacent lymph node (+) Metastasis (-) Metastasis (+)
Cases 30 15 18 18 15 23 10
CR1 (A405) 1.252±0.31 0.95±0.22a 0.81±0.29ab 0.88±0.27a 0.73±0.25ab 0.78±0.26a 0.44±0.29ad
Adhering activity (%) 64.2±7.4 52.8±6.7a 48.1±7.2ab 47.6±7.0a 41.1±7.1ab 46.1±6.1a 38.9±6.2ad

bP<0.01 vs normal control; aP<0.05, dP<0.01 vs negative.

DISCUSSION
The immune functions of erythrocytes have been studied at gene levels[5-8]. RBCs are the major cellular component of the peripheral blood and occupy 50% of the total blood volume[9]. Normal RBC cytosol contains the majority of natural killer enhancing factors and other factors, such as PIF. Damaged RBC could produce tumor necrosis factor inducing factor. RBCs appear to have an important regulating role in immune function, and RBC activity involves in regulating multiple cytokines such as IL-2, IL-3, CSF, IL-6, TNF-r, TNF. CD44 and CD58 expressed by RBC could serve as the center in controlling immune status, which can directly affect the development and progression of tumors[8-11]. The changes of CR1 quantitative expression and adhering activity are consanguineously related to the development and metastasis of liver carcinoma.
      In our study, the number and adhering activity of CR1 high expression genomic type gallbladder carcinomas were significantly lower than those in healthy individuals, indicating that defective expression of CR1 in gallbladder carcinoma is mostly acquired through central peripheral mechanisms.
      Gallbladder carcinoma is often associated with cholecyst-olithiasis and cholecystitis in 40-100% of cases[19-28]. Although no carcinogenic substance has so far been isolated from the bile or the stones in patients with cholecystolithiasis and cholecystitis, many scholars suggested that gallstone might play a role as a chronic injury factor to induce a series of epithelial pathologic changes[29-38]. Gong et al.[17] reported that in 150 consecutive cholecystectomy specimens for detection of cholelithiasis or cholecystitis, 76.68% exhibited epithelial hyperplasia, 16.89% atypical hyperplasia, 1.32% carcinoma in situ and 2.11% invasive carcinoma. Simple epithelial hyperplasia was found in the mucus adjacent to invasive carcinoma.With the passage of time, a significant number of atypical hyperplasiae presumably would progress to a higher grade lesion, becoming carcinoma. Toyonaga et al.[39] reported that in 200 consecutive cholecystectomy specimens for detection of cholelithiasis or cholecystitis, 83% simple epithelial hyperplasiae, 13.5% atypical hyperplasiae and 3.5% carcinomas in situ. In general, a significant number of atypical hyperplasiae presumably would progress to a higher grade lesion, 80% atypical hyperplasiae could become pre-cancer lesions. Albores-Saavedra et al.[11] results showed that cholelithiasis or cholecystitis produced a series of epithelial pathologic changes, such as simple epithelial hyperplasia, atypical hyperplasia and carcinoma in situ, which represented the precancer lesion of gallbladder carcinoma. The probable sequence of events appear to be as follows. Hyperplasia has been found to have atypical hyperplasia which in turn may progress to neoplasia. With the passage of time, a significant number of atypical hyperplasiae presumably would progress to carcinoma in situ and invasive carcinoma[24]. So almost all scholars have suggested that a small number of hyperplasiae of the gallbladder would evolve toward in situ carcinoma which finally becomes invasive carcinoma[40]. In our study, the number and adhering activity of CR1 in patients with gallbladder carcinoma were significantly lower than those in patients with chronic cholecystitis and cholelithiasis and healthy individuals (P<0.01). The number and adhering activity of CR1 in patients with chronic cholecystitis and cholelithiasis were significantly lower than those in healthy individuals. There was a positive linear correlation between quantitative expression and the adhering activity of ECR1 (r = 0.79, P<0.01). Compared with data the on preoperative day, the number and adhering activity of CR1 in patients with gallbladder carcinoma decreased greatly on the third postoperative day and  in the first postoperative week, indicting that surgery plays an injurious role in the disorder of RBC and cytokine functions[12,41,42]. The number and adhering activity of CR1 recovered in the second postoperative week, and increased greatly in the third postoperative week.
     The number and the adhering activity of CR1 in gallbladder carcinoma patients with infiltrating, adjacent lymph node and distant organ metastases were significantly lower than those in patients without metastasis. This study demonstrates CR1 can be used as an immune therapy for gallbladder carcinoma.

REFERENCES
1    Jiao XY, Lü MD, Huang JF. Current advances in the risk factors for gallbladder carcinoma. Zhonghua Putong Waike 
      Zazhi 2002; 17: 117-119
2    L
ü MD, Jiao XY. Current status of research on tumor markers of biliary tumors. Zhongguo Shiyong Waike Zazhi 
      2001; 9: 522-523
3    Jiao XY, Li DM, L
ü MD, Huang JF, Liang LJ. Significance of nucleolar organizers regions associated proteins of T
      lymphocytes in diagnosis and monitoring of gallbladder carcinoma. Zhonghua Putong Waike Zazhi 
      2002; 117: 313-314
4    Zou SQ, Zhang L. Relative risk factors analysis of 3922 cases of gallbladder cancer. Zhonghua Waike Zazhi 
      2000; 38: 800-805
5    Birmingham DJ, Chen W, Liang G, Schmitt HC, Gavit K, Nagaraja HN. A CR1 polymorphism associated with constitutive
      erythrocyte CR1 levels affects binding to C4b but not C3b.Immunology 2003; 108: 531-538
6    Rowe JA, Raza A, Diallo DA, Baby M, Poudiougo B, Coulibaly D, Cockburn IA, Middleton J, Lyke KE, Plowe CV, 
      Doumbo OK, Moulds JM. Erythrocyte CR1 expression level does not correlate with a Hind III restriction fragment length
      polymorphism in africans; implications for studies on malaria susceptibility. Genes Immun 2002; 3: 497-500
7    Zorzetto M, Bombieri C, Ferrarotti I, Medaglia S, Agostini C, Tinelli C, Malerba G, Carrabino N, Beretta A, Casali L, 
      Pozzi E, Pignatti PF, Semenzato G, Cuccia MC, Luisetti M. Complement receptor 1 gene polymorphisms in sarcoidosis. 
      Am J Respir Cell Mol Biol 2002; 27: 17-23
8    Nagayasu E, Ito M, Akaki M, Nakano Y, Kimura M, Looareesuwan S, Aikawa M. CR1 density polymorphism on 
      erythrocytes of falciparum malaria patients in Thailand. Am J Trop Med Hyg 2001; 64: 1-5
9    Moulds JM, Zimmerman PA, Doumbo OK, Kassambara L, Sagara I, Diallo DA, Atkinson JP, Krych-Goldberg M, Hauhart RE,
      Hourcade DE, McNamara DT, Birmingham DJ, Rowe JA, Moulds JJ, Miller LH. Molecular identification of knops blood 
      group polymorphisms found in long homologous region D of complement receptor 1. Blood 2001; 97: 2879-2885
10  Wang HB, Qian BH, Niu F, Jiang AQ. Changes of genomic density polymorphism and quantitative expression of 
      complement receptor type 1 in patients with liver diseases. Zhonghua Ganzangbing Zazhi 2000; 8: 335-337
11  Wang HB, Zhao XP, Guo F. Changes of quantitative expression, adhering activity and genomic density polymorphism of
      complement type1 in erythocytes (CR1) of patients with liver cancer. Zhonghua Jianyan Yixue Zazhi 20001; 24: 31-33
12  Jiao XY, Shi JS, Wang JS, Yang JY, He P. Effects of radical cholecystectomy on nutritional and immune status in patients
      with gallbladder carcinoma. World J Gastroenterol 2000; 6: 445-447
13  Jiao XY, Shi JS, Gao JS, Zhou LS. Determination of levels of cellular immunity and humoral immunity in patients with
      gallbladder carcinoma. Zhongguo Puwai Jichu Yu Linchuang Zazhi 1999; 4: 227-229
14  Jiao XY, Shi JS, Gao JS, Zhou LS, Han WS, Liu G, Lu Y. Study on the serum IL-2, SIL-2r and CEA levels in patients with
      gallbladder carcinoma. Zhonghua Gandan Waike Zazhi 1999; 5: 342
15  Shi JS, Wang JS, Liu G, Yu YL, Lu Y, Jiao XY, Yang YJ, Li GC, Han Y. Early diagnosis of primary gallbladder carcinoma.
      Hepatobiliary Pancreat Dis Int 2002; 1: 273-275
16  Nevin JE, Moran TJ, Kay S, King R. Carcinoma of the gallbladder: staging, treatment, and prognosis. Cancer 
      1976; 37: 141-148
17  Gong FQ, Gao ZY, Wang JB, Liu SG, Liu JQ. Pathologic study of precancerous lesion of the gallbladder. Zhonghua 
      Zhongliu Zazhi 1989; 11: 127-129
18  Cornillet P, Philbert F, Kazatchkine MD, Cohen JHM. Genomic determination of the CR1 (CD35) density polymorphism 
      on erythrocytes using polymerase chain reaction amplification and Hind III restriction enzyme digestion. J Immunol
      Methods 1991; 136: 193-197
19  Noshiro H, Chijiiwa K, Yamaguchi K, Shimizu S, Sugitani A, Tanaka M. Factors affecting surgical outcome for
      gallbladder-carcinoma. Hepatogastroenterology 2003; 50: 939-544
20  Wakai T, Shirai Y, Yokoyama N, Ajioka Y, Watanabe H, Hatakeyama K. Depth of subserosal invasion predicts long-term
      survival after resection in patients with T2 gallbladder carcinoma. Ann Surg Oncol 2003; 10: 447-454
21  Ishikawa T, Horimi T, Shima Y, Okabayashi T, Nishioka Y, Hamada M, Ichikawa J, Tsuji A, Takamatsu M, Morita S.
      Evaluation of aggressive surgical treatment for advanced carcinoma of the gallbladder. J Hepatobiliary Pancreat Surg
      2003;  10: 233-238
22  Nakakubo Y, Miyamoto M, Cho Y, Hida Y, Oshikiri T, Suzuoki M, Hiraoka K, Itoh T, Kondo S, Katoh H. Clinical significance 
      of immune cell infiltration within gallbladder cancer. Br J Cancer 2003; 89: 1736-1742
23  Enomoto T, Todoroki T, Koike N, Kawamoto T, Matsumoto H. Xanthogranulomatous cholecystitis mimicking stage IV 
      gallbladder cancer. Hepatogastroenterology 2003; 50: 1255-1258
24  Jarnagin WR, Ruo L, Little SA, Klimstra D, D’Angelica M, DeMatteo RP, Wagman R, Blumgart LH, Fong Y. Patterns of 
      initial disease recurrence after resection of gallbladder carcinoma and hilar cholangiocarcinoma: implications for 
      adjuvant therapeutic strategies. Cancer 2003; 98: 1689-1700
25  Yanagisawa N, Mikami T, Yamashita K, Okayasu I. Microsatellite instability in chronic cholecystitis is indicative of an 
      early stage in gallbladder carcinogenesis. Am J Clin Pathol 2003; 120: 413-417
26  Shukla VK, Adukia TK, Singh SP, Mishra CP, Mishra RN. Micronutrients, antioxidants, and carcinoma of the gallbladder. 
      J Surg Oncol 2003; 84: 31-35
27  Kokudo N, Makuuchi M, Natori T, Sakamoto Y, Yamamoto J, Seki M, Noie T, Sugawara Y, Imamura H, Asahara S, 
      Ikari T. Strategies for surgical treatment of gallbladder carcinoma based on information available before resection. 
      Arch Surg 2003; 138: 741-750
28  Bani-Hani KE, Yaghan RJ, Matalka II, Shatnawi NJ. Gallbladder cancer in northern Jordan. J Gastroenterol Hepatol 
      2003; 18:954-959
29  Malik IA. Clinicopathological features and management of gallbladder cancer in pakistan: A prospective study of 233
      cases. J Gastroenterol Hepatol 2003; 18: 950-953
30  Andrea C, Francesco C. Squamous-cell and non-squamous-cell carcinomas of the gallbladder have different risk factors.
      Lancet Oncol 2003; 4: 393-394
31  Hemminki K, Li X. Familial liver and gall bladder cancer: a nationwide epidemiological study from Sweden. Gut 
      2003; 52: 592-596
32  Kondo S, Nimura Y, Kamiya J, Nagino M, Kanai M, Uesaka K, Yuasa N, Sano T, Hayakawa N. Factors influencing
      postoperative hospital mortality and long-term survival after radical resection for stage IV gallbladder carcinoma. World 
      J Surg 2003;27: 272-277
33  Haratake J, Kasai T, Makino H. Diffuse mucosal carcinoma of intrahepatic and extrahepatic bile ducts including
      gallbladder. Pathol Int 2002; 52: 784-788
34  Pandey M. Risk factors for gallbladder cancer: a reappraisal. Eur J Cancer Prev 2003; 12: 15-24
35  Shi JS, Zhou LS, Jiao XY, Lu Y, Hao XY, Han WS, Liu G. Clinical significance of detecting tumor necrosis factor and its
      receptor in serum and bile in patients with gallbladder cancer and gallstone. Zhonghua Gandan Waike Zazhi 
      2001; 7: 82-83
36  Misra S, Chaturvedi A, Misra NC, Sharma ID. Carcinoma of the gallbladder. Lancet Oncol 2003; 4: 167-176
37  Parwani AV, Geradts J, Caspers E, Offerhaus GJ, Yeo CJ, Cameron JL, Klimstra DS, Maitra A, Hruban RH, Argani P.
      Immunohistochemical and genetic analysis of non-small cell and small cell gallbladder carcinoma and their precursor
      lesions. Mod Pathol 2003; 16: 299-308
38  Rashid A. Cellular and molecular biology of biliary tract cancers. Surg Oncol Clin N Am 2002; 11: 995-1009
39  Toyonaga T, Chijiiwa K, Nakano K, Noshiro H, Yamaguchi K, Sada M, Terasaka R, Konomi K, Nishikata F, Tanaka M.
      Completion radical surgery after cholecystectomy for accidentally undiagnosed gallbladder carcinoma. World J Surg
      2003;27: 266-271
40  Henson DE, Albores-Saavedra J, Corle D. Carcinoma of the gallbladder carcinoma, histologic types, stage of disease,
      grade, and survival rates. Cancer 1992; 70: 1493-1497
41  Albores-Saavedra J, Alcantra-Vazquez A, Cruz-Ortiz H, Herrera-Goepfert R. The precursor lesions, of invasive 
      gallbladder carcinoma, hyperplasia, atypical hyperplasia and carcinoma in situ. Cancer 1980; 45: 919-927
42  Fan YZ, Zhang JT, Yang HC, Yang YQ. Expression of MMP-2,TIMP-2 protein and the ratio of MMP-2/TIMP-2 in 
      gallbladder carcinoma and their significance. World J Gastroenterol 2002; 8: 1138-1143

   Edited by Wang XL and Ren SY  Proofread by Xu FM 

 

Reviews Add
more>>


Related Articles:
Expression of MMP-2,TIMP-2 protein and the ratio of MMP-2/TIMP-2 in gallbladder carcinoma and their significance
Metachronous bile duct cancer nine years after resection of gallbladder cancer
Xanthogranulomatous cholecystitis mimicking gallbladder carcinoma with a false-positive result on fluorodeoxyglucose PET
Genetic changes of p53, K-ras, and microsatellite instability in gallbladder carcinoma in high-incidence areas of Japan and Hungary
A clinicopathological analysis in unsuspected gallbladder carcinoma: A report of 23 cases
more>>