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Background: Although the standard 3-week capecitabine regimen (1250 mg/m 2 twice daily for 2 weeks followed by a 1-week rest) has shown superior activity and improved safety over bolus 5-fluorouracil/leucovorin in two large randomized phase III trials in Europe and in the United States, only a 4-week regimen of capecitabine (828 mg/m 2 twice daily for 3 weeks) has been studied in Japan. Therefore, we performed a phase II study to investigate the 3-week regimen of capecitabine in Japanese patients with metastatic colorectal cancer (MCRC). Methods: Previously untreated patients with MCRC received oral capecitabine 1250 mg/m 2 twice daily for 2 weeks. Treatment was repeated every 3 weeks. Blood and urine samples were collected for pharmacokinetic analysis. Results: Sixty patients were enrolled. The overall response rate was 35% [95% confidence interval (CI), 23–48%], and 52% of patients had stable disease. The median time to progression was 5.5 months (95% CI, 4.2–6.7 months). The median overall survival was 20.2 months (95% CI, 16.6–27.8 months). The most frequently occurring adverse drug reaction was hand-foot syndrome (all-grade 73%; grade 3 13%). Diarrhea, anorexia, nausea and stomatitis were each seen in 37% of patients. The pharmacokinetic profiles of capecitabine and its metabolites were similar to those reported in Caucasian patients. Conclusions: The 3-week regimen of capecitabine was effective and well tolerated in Japanese patients with MCRC as well, and could be used as the basic regimen for future combination therapies.

症例1は50歳の女性で, 直腸癌の診断で当院紹介された. 直腸Raに2型病変を認め, CT上傍大動脈リンパ節腫大が見られた. 低位前方切除術施行し, 術中迅速組織診で傍大動脈リンパ節転移陽性であった. 術後化学療法を1クール施行し, 傍大動脈リンパ節腫大は消失したが術後4年9か月のCTで再び傍大動脈リンパ節転移再発を認めた. 化学療法4クール施行後, リンパ節腫大は消失した. 術後8年9か月時点で再発なく経過中である. 症例2は29歳の男性で, 下血にて前医入院し, 精査加療目的で当院転院した. 歯状線に接して粘膜下腫瘍様隆起を認め, 生検で小細胞癌の診断となった. 化学療法3クールと放射線照射を施行し腫瘍の縮小を得た後, 腹会陰式直腸切断術を施行した. 原発巣, リンパ節共に腫瘍細胞を認めず, 加療後19か月現在無再発生存中である. 直腸原発小細胞癌は予後不良だが集学的治療が著効する例もあることから積極的な治療が望まれる.

症例は75歳の女性で, 1997年1月にS状結腸癌に対して切除術を受けた (well, se, ly1, v0, n0; Stage II). 術後, 追加治療せず外来経過観察としたが, 2002年11月より血清CEA値の上昇を認め, 2003年3月の腹部CTで脾臓に4×2cm大の腫瘤像が確認された. 全身F-18 fluorodeoxyglucose positron emission tomographyでは同部に強い集積が認められたため, 脾臓転移を疑い2003年5月脾摘術を施行した. 病理組織学的に前回切除されたS状結腸癌の脾臓転移と診断した. その後も補助化学療法を行わず, 術後19か月経過した現在, 再発なく生存中である. 大腸癌の脾臓への孤立性転移はまれであり, 若干の文献的考察を加えて報告する.

目的:大腸癌に対する腹腔鏡補助下大腸切除手術(LAC)と開腹手術(OPEN)のリンパ節郭清が同等の質であるかをリンパ節の採取個数で比較検討した.対象と方法:右半結腸切除,S状結腸切除,直腸高位前方切除が施行された大腸癌手術症例77例.リンパ節採取個数を各リンパ節領域ごとに比較検討した.結果:右半結腸切除21例(LAC:OPEN;12例:9例),S状結腸切除41例(26例:15例),前方切除15例(8例:7例)であった.腫瘍径による補正後の摘出リンパ節個数は(以下LAC:OPEN),右半結腸切除;1群(15.4±1.3:17.5±3.6),主幹2群(8.7±1.0:8.2±0.7),壁在2群(10.8±1.6:15.3±2.9),主幹3群(7.5±1.0:7.6±0.8),S状結腸切除;1群(12.3±1.3:13.1±1.5),主幹2群(8.2±1.2:8.1±1.2),壁在2群(7.0±1.4:5.7±0.9),主幹3群(6.8±1.0:6.4±1.3),前方切除;1群(12.5±2.3:12.4±2.7),主幹2群(9.3±1.4:9.7±2.3),壁在2群(6.8±2.4:5.6±1.3),主幹3群(5.0±0.9:5.3±1.3)であった.考察:LACとOPENで以上の3種類の術式においてリンパ節郭清はほぼ同等であることが示唆された., Aim: This study was conducted to evaluate. The number of dissected lymph nodes between laparoscopyassisted colectomy (LAC) and open colectomy (OPEN). Methods: Seventy-seven patients who underwent hemi-colectomy (LAC: OPEN; 12cases: 9cases), sigmoidectomy (LAC: OPEN; 26cases: 15cases) and high anterior resection (LAC: OPEN; 8cases: 7cases) at Kanagawa Cancer Center were entered in this study. The number of dissected lymph nodes at each N1, paracolic N2, central N2, and central N3 area was compared. Results: In Hemi-colectomy, N1 area was LAC: OPEN =15.4±1.3 : 17.5±3.6 (p = 0.548), Central N2 was 8.8±1.0 : 8.2±0.7 (p = 0.684) , Paracolic N2 was 10.8±1.6 : 15.3±2.9 (p = 0.176), and Central N3 was 7.5±1.0 : 7.6 ± 0.8 (p = 0.966) . In sigmoidectomy, Nl was 12.3±1.3 : 13.1±1.5 (p = 0.675) , Central N2 was 8.2±1.2 : 8.1±1.2 (p = 0.945) , Paracolic N2 was 7.0±1.4 : 5.7±0.9 (p = 0.522) , Central N3 was 6.8±1.0 : 6.4±1.3 (p = 0.808) . In Anterior resection, N1 was 12.5±2.3 : 12.4±2.7 (p = 0.969) , Central N2 was 9.3±1.4 : 9.7±2.3 (p = 0.863) , Paracolic N2 was 6.8±2.4 : 5.6±1.3 (p = 0.689) , and Central N3 was 5.0±0.9 : 5.3±1.3 (p=0.879). Conclusion: The results suggested that quality of lymph node dissection in LAC was equal to that of OPEN.

はじめに: 大腸癌根治手術後の早期再発発見を目的としたintensive follow up法の妥当性を検討した. 対象および方法: 1986～2001年までの大腸癌根治度A症例のうち単発再発を認めた126例を対象とした. 4か月に1回のCTにより再発を発見された症例をintensive群(以下, I群, n=66), 1年に1 回のCTにより再発を発見された症例をconventional群 (以下, C群, n=60) に分類し, 生存期間 (ST), 無再発期間 (DFI), 再発後生存日数 (SAR) をretrospectiveに検討した. 結果: DFIはI群が467日, C群が690日で有意差を認めたがSARはそれぞれ943日, 767日であり有意差はなかった. また臓器別に検討すると肺再発のSARはI 群で1,296日, C 群で925日であり有意差はないものの改善される傾向を認めた. 考察: CTによる大腸癌術後intensive follow upは再発後生存日数の向上には寄与しなかった. しかし腫瘍マーカー, 問診でスクリーニングが困難な肺再発においては再発後生存日数が向上する可能性が示唆された.

症例は69歳の男性.平成15年6月6日から,他院で診断された左肺癌に対して放射線化学療法を施行していたが, 6月24日ごろより粘血便が出現した.腹部に所見はみられず,大腸内視鏡検査で直腸Rs内に球状の腫瘤を認め,生検では壊死組織であった.腸重積発症の恐れがあるため,平成15年8月22日にS状結腸切除術を施行した.手術所見ではS状結腸が直腸Rs内に陥入して重積様の所見を呈していた.切除標本ではS状結腸内腔に著明に突出した球状腫瘍を認め,その口側に2型進行癌が隣接して存在していた.球状腫瘤は組織学的に紡錐形細胞が索状配列しており,隣接するS状結腸浸潤癌近傍の粘膜下から発生し,これを押し上げていると考えられた.免疫染色ではc-kit, CD34, SMA, S100ともに陰性であり広義のGISTと診断された.結腸GISTの報告例は20例と少なく,さらにこのような肉眼形態は非常に稀であるため,本邦報告例の集計とともに報告する.

We examined whether sustained alleviation of inflammation as monitored by serum alanine aminotransferase (ALT) levels was associated with longer survival in hepatectomized hepatocellular carcinoma (HCC) patients with hepatitis C virus-associated liver cirrhosis (HCV-LC). Thirty-four hepatectomized patients with HCV-LC and HCC as a single nodule, and for whom more than 5 years had elapsed after the hepatectomy, were studied. They had no histologic evidence of portal or hepatic vein invasion. They were subdivided into two groups according to their serum ALT levels in the 2 years after hepatectomy: the low ALT group comprised 13 patients whose serum ALT levels showed a sustained low level below 80 IU, and the high ALT group comprised 21 patients whose serum ALT levels showed several peaks or plateaus above 80 IU. The patients had been followed-up prospectively with frequent ultrasonography and magnetic resonance imaging or computed tomography for recurrence for > 5 years. The survival period, non-recurrence interval and number of recurrences were observed. Recurrences were treated with transcatheter chemoembolization in all cases. The cumulative survival rate in the low ALT group was significantly better than that in the high ALT group (P < 0.05). The 5-year survival in the low ALT group was as high as 92.3% (12 of 13) compared with 33.3% (7 of 21) in the high ALT group (P < 0.05). The cumulative non-recurrence rate in the low ALT group was also significantly better than that in the high ALT group (P < 0.01). The survival period correlated well with the interval until the first recurrence (r = 0.545, P = 0.006). There was a tendency for the number of recurrences in the low ALT group (1.5 +/- 0.4, mean +/- SE) to be fewer than that in the high ALT group (2.2 +/- 0.4), although this was not significant. Sustained alleviation of inflammation, as indicated by low ALT levels, provides a survival advantage mainly due to the longer non-recurrence interval, and possibly because of fewer recurrences, in hepatectomized HCC patients with HCV-LC.

目的: 大腸癌におけるリンパ節転移個数の予後予測における有用性を検討する.対象: 1990年から2000年までに治癒切除された大腸癌 (肛門管癌除く) 患者815例 (男性487例女性328例, 62.7歳). stage 0 33例, stage I 174例, stage II 220例, stage IIIa 192例, stage IIIb 136例, stage IV 60例. リンパ節転移個数は0個, 1～3個 (1個, 2～3個), 4個以上の3群 (4群) に分け, n因子は大腸癌取扱い規約に従い検討した.結果: 術後生存日数において単変量解析でn因子はn2, n3, n4の各群間に有意差はみられなかった. リンパ節転移個数は各群間に有意差をみた. 深達度, 肝転移, 腹膜播種, 遠隔転移を加えた多変量解析は両者有意な予後規定因子となったが, n因子のrelative riskはn3とn4の間で逆転したのに対し転移個数は1次関数的に増加した. 無再発日数においても単変量解析で転移個数はすべての群で有意差がみられたが, n因子はn2, n3, n4の間に有意差はみられなかった. 多変量解析ではn因子が有意ではあるがrelative riskはn4群がn3群に比べ低値 (n3 6.760, n4 5.722) となったのに対し, リンパ節転移個数ではすべて1次関数的に増加した.結語: リンパ節転移個数は生存率再発予測について, 単変量でも多変量解析でもn因子より有用である. リンパ節転移についてはその部位と共に転移個数についても考慮すべきであり, これらを術中所見の判定に用いることにより, 適切な大腸癌のリンパ節郭清の一助となると考えられた.

症例は50歳の女性. 1996年6月注腸検査, 大腸内視鏡検査で肛門縁から10cmの直腸に2型病変をみとめ生検でanaplastic squamous cell carcinomaであった. 術前CTにて傍直腸, 大動脈周囲, 左腸骨動脈周囲にリンパ節腫大を認めた. 8月12日低位前方切除術施行, 術中迅速診にて大動脈周囲リンパ節に転移陽性であり, 根治度Cであった. 病理組織診では直腸原発の小細胞癌と診断した. 肺小細胞癌に準じ術後CDDP40mg×2日, VP-16 100mg×3日(1クール)の化学療法を施行した. 化学療法が奏効し術後9MのCTにて傍大動脈リンパ節腫大消失CRを得て以後2001年2月まで42か月, 再発なく経過中である. 小細胞癌の直腸原発例はまれであり, 1クールにもかかわらず化学療法が奏効した1例を経験したので報告する.

We investigated whether or not a high serum alanine aminotransferase (ALT) level is associated with a more rapid recurrence of hepatocellular carcinoma (HCC) in hepatectomized patients with hepatitis C virus (HCV)-associated liver cirrhosis (LC) (HCV-LC) and HCC. Thirty-three hepatectomized patients with HCV-LC and HCC of a single nodule who had no histologic evidence of portal or hepatic vein invasion and who had been followed up for more than 3 years were included in the study. They were subdivided into two groups according to their serum ALT levels, ALT being a well-known marker of inflammatory necrosis in the liver. Seventeen patients whose serum ALT levels showed several peaks or plateaus above 80 international units (IU) were designated as the high ALT group, and 16 patients whose serum ALT levels showed a sustained low level below 80 IU until the first recurrence were designated as the low ALT group, and the interval between hepatectomy and the first recurrence was observed. In the high ALT group, HCC recurred within 3 years in 70.6% of the patients. In contrast, it recurred in only 18.8% of the low ALT group within the same period (p < 0.05). There was a significant difference (p = 0.0201) between the two groups in the cumulative nonrecurrence rate. The mean interval in recurrent patients between hepatectomy and the first recurrence in the high ALT group (23.6 ± 2.8 months; mean ± SE) was significantly (p < 0.02) shorter than that in the low ALT group (49.3 ± 9.7 months). The expected interval between hepatectomy and recurrence was as short as 2.8 ± 0.5 years (mean ± SE) in the high ALT group, compared with 5.8 ± 0.7 years in the low ALT group (p < 0.05). These results showed that the recurrence of HCC was accelerated in the high ALT group, suggesting that suppression of the rise in ALT level after hepatectomy by treatment with anti-inflammatory drugs may prolong the interval until recurrence by about 2 years in hepatectomized patients with HCC and HCV-LC.

A total of 32 patients, who had undergone surgical currative resection of colorectal cancer liver metastasis, were retrospectively analyzed. Recurrences were observed in 75% of the 32 patients. These recurrences were hepatic in 33.3% of patients with recurrence, intra-and extrahepatic in 37.5% and extrahepatic in 29.2%. The residual liver recurrence rate was significantly higher in H2H3 cases than in H1 cases. The five year survival rate of H1 cases was 50.0%, while none of the H2H3 patients survived more than three years. Advanced wall invasion in the primary tumor was unfavorable prognostic indicator in H1 cases, without statistical significance. The patients who underwent adjuvant intrahepatic arterial chemotherapy had a favorable residual liver recurrence rate and survival rate, but this did nor reach statistical significance. To prolong survival in patients with undetected extrahepatic residual disease at the time of hepatic resection, effective adjuvant therapy, combining hepatic arterial chemotherapy and systemic chemotherapy, is required.

To examine whether the marked increase in DNA synthesis of hepatocytes in cirrhotic liver might elicit multicentric hepatocarcinogenesis, we examined the relationship between new development of hepatocellular carcinoma (HCC) and the bromodeoxyuridine (BrdU) labeling index (LI) of hepatocytes in the residual liver of hepatectomized patients with liver cirrhosis (LC) and HCC. Eighteen hepatectomized patients with LC and HCC, whose resected liver revealed neither portal nor hepatic vein invasion by histologic examination, were studied (to exclude cases with intrahepatic metastasis). DNA synthesis activity of hepatocytes from the residual cirrhotic liver was measured by a BrdU/anti-BrdU in vitro method. The incidence of HCC recurrence was studied during a 3-year follow-up period. Among 18 patients, 9 patients had recurrence and 9 did not. The average BrdU LI in the recurrent patients was 2.6 +/- 1.3% and was significantly higher than that in patients without recurrence (1.4 +/- 0.5%, P < 0.05). All five patients who had a BrdU LI of 2.4% or above showed recurrence within 3 years, as compared with 4 of 13 (30.8%) patients with BrdU LI of less than 2.4% (P < 0.05). Our data indicate that abnormally high DNA synthesis in hepatocytes in the background cirrhosis might lead to the development of multicentric carcinogenesis in human cirrhotic liver, and in the residual cirrhotic liver of hepatectomized patients with HCC and LC, it may be a predictor of new development of HCC.

BACKGROUND There is a concept that a cancer often maintains some of the traits of the background tissue cells. Thus, the possibility exists that the DNA synthetic activity of the hepatocytes in cirrhotic tissue affects that of hepatocellular carcinoma (HCC) cells. METHODS DNA synthesis activity of hepatocytes from background cirrhotic tissue and HCC cells from cancerous tissue was studied in 30 patients with liver cirrhosis (LC) and HCC who had undergone hepatectomy; the study was done using the bromodeoxyuridine (BrdU)-anti-BrdU in vitro method. The correlation between the BrdU labeling index (LI) of hepatocytes from noncancerous cirrhotic tissue and that of HCC cells was studied. The relationship between both BrdU LI values and the survival after hepatectomy also was studied. RESULTS A significant correlation (r = 0.572; P < 0.001) was identified between the BrdU LI for HCC and that for noncancerous cirrhotic tissue. A significant correlation (r = -0.572; P < 0.05) was found between the BrdU LI of HCC cells and the survival after hepatectomy. A significant correlation (r = -0.678; P < 0.01) was observed between the BrdU LI of the noncancerous cirrhotic tissue and the survival. CONCLUSIONS It was concluded that a significant correlation existed between DNA synthesis of hepatocytes from the background cirrhotic tissue and that of HCC cells and that DNA synthesis activity of hepatocytes from noncancerous cirrhotic tissue and that of HCC cells from cancerous tissue could predict the survival of patients with HCC who had undergone hepatectomy.

Bromodeoxyuridine (BrdU) labeling indices (LI) of liver biopsied specimens from 27 control liver cirrhosis patients (control LC) without hepatocellular carcinoma (HCC), 26 cirrhotic patients with HCC, and from 6 control subjects were examined using an in vitro BrdU labeling technique. The mean BrdU LI +/- S.E. of HCC cancerous portion, HCC non-cancerous cirrhotic portion, control LC, and control subjects were 6.6 +/- 0.8%, 2.7 +/- 0.3%, 1.7 +/- 0.3% and 0.25 +/- 0.09%, respectively. Interestingly, there was a significant difference (p less than 0.025) between the non-cancerous LC portion and control LC. While 25 of 26 non-cancerous LC portion of HCC patients were in the high LI group (greater than or equal to 1.4%), 15 of 27 in the control LC were in the low LI group (less than 1.4%) (p less than 0.001). Of the 12 control LC with high LI, 6 developed HCC within 2 years, whereas only one of the 15 control LC with low LI developed HCC (p less than 0.05). In conclusion, HCC may develop in cirrhotic patients with high DNA synthetic potency.

Tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in plasma has been reported to be related to disease progression in patients with colorectal cancer. However, the prognostic significance of plasma TIMP-1 has not been clarified.Concentrations of TIMP-1 protein were measured by enzyme-linked immunosorbent assay in plasma samples of 87 preoperative patients who subsequently underwent resection, and prognosis was compared. The cut-off value of plasma TIMP-1 was defined as 170 ng/ml.When clinicopathological factors between patients with positive and those with negative plasma TIMP-1 were analyzed, significant differences were observed in lymph node metastasis, serosal invasion, curability and Dukes' classification. Univariate analysis of these factors demonstrated that depth of invasion, metastases to lymph nodes, peritoneum, liver and distant organ, lymphatic and vessel invasions, curability. Dukes' classification and plasma TIMP-1 concentration were significant. By multivariate analysis excluding patients with distant or peritoneal metastases, histological type, lymphatic invasions, lymph node metastasis and plasma TIMP-1 were retained in the final model.These results suggested that plasma TIMP-1 may be a useful prognostic marker for survival in patients with colorectal carcinoma.

The relationship between the recurrence of hepatocellular carcinoma (HCC) and the serum alanine aminotransferase (ALT) level was studied in hepatectomized patients with hepatitis C virus (HCV)-associated cirrhosis and HCC.Twenty-six hepatectomized patients with HCV-associated cirrhosis and HCC whose resected specimens showed neither portal vein nor hepatic vein invasion by HCC histologically were divided into 2 groups: 15 patients who had no recurrence 3 years after surgery (Group A) and 11 patients whose disease recurred 1-3 years after surgery (Group B). The patients' serum ALT levels during this period were examined.In Group A, serum ALT generally showed sustained low levels80 international units (INU) in 12 patients (80%). In contrast, ALT levels in Group B showed several peaks or plateaus80 INU in all patients except 2. The recurrence rate of HCC in the hepatectomized patients with sustained low levels of ALT was 14.3% (2 of 14 patients) at 3 years, and was significantly lower (P0.01) than that in those patients whose ALT levels showed several peaks or plateaus80 INU (9 of 12 patients; 75.0%). The average level of mode of ALT in Group A (48.8 +/- 26.0 INU) was significantly smaller than that in Group B (101.1 +/- 47.3 INU) (P0.005).The importance of hepatocytic necrosis in the recurrence of HCC in hepatectomized patients with cirrhosis and HCC of HCV origin was demonstrated and the significance of subsiding hepatic necroinflammatory process in the prevention of HCC recurrence suggested.

To clarify sex differences in DNA synthesis in hepatocellular carcinoma (HCC), bromodeoxyuridine labeling indices (BrdU LI) of HCC cells included in tumor biopsy specimens from 12 consecutive male patients and from 5 consecutive female patients all with liver cirrhosis and HCC were examined using an in vitro labeling technique. The mean BrdU LI +/- SE of HCC from male patients (7.7 +/- 0.8%) was significantly (P less than 0.05) higher than that from female patients (4.4 +/- 1.0%). While 7 of the 12 male HCC patients belonged to the high DNA synthesis group (BrdU LI greater than or equal to 7.0%), none of the 5 female HCC patients showed high DNA synthesis (P less than 0.05). We conclude that DNA synthesis in HCC was higher in males than in females.

ThymidineのanalogueであるBrdUに対するモノクローナル抗体を用いて肝細胞癌(以下HCC),肝硬変(以下LC),慢性活動性肝炎(以下CAH),アルコール性肝線維症(以下Al-F)等の各種肝疾患時の肝のDNA合成を免疫組織学的に検討した.即ち,上記疾患の肝生検組織をBrdUを含んだ培養液とincubationしBrdU摂取をin vitroで測定した.取り込みが最も高率であったのはHCC癌部でL.I.は5.7±0.9%であり,次いでHCC非癌硬変部(3.4±0.4%),さらにHCCの無いLC (2.3±0.7%), Al-F (2.4±0.7%)であった.一方,CAHでは1.0±0.1%と予想外に低かった.HCC癌部は,HCC非癌硬変部(p

ThymidineのanalogueであるBromodeoxyuridine(以下BrdU)に対するモノクロナール抗体を用いて原発性肝癌(以下HCC) 16例の腫瘍細胞のDNA合成を免疫組織学的に検討した.方法は0.1%にBrdUを含んだRPMI1640液と肝生検組織をincubationし,BrdU摂取率をin vitroで測定した.Labeling Index (L.I.)のMean±SEは7.06±2.71%で症例間で2.6%から12.7%と大きな差異がみられた.又,HCCのL.I.は他の消化器癌に比べて概して低値であった.次にHCCの組織学的差異によるBrdU L.I.の検討ではL.I. 5%未満では全てtrabecular typeであったのに対し,5%以上ではpseudoglandular typeやsolid typeが高頻度にmixしていた.一方,Edmondson & Steiner'sの異型度分類とBrdU L.I.との関係では,より異型度の高いIII型がL.I. 10.64±2.10%で,I～II型の5.87±1.61%に比べ有意に高く,異型度とDNA合成能との相関が示唆された.

For anticancer effects on metastatic lymph node, activated carbon adsorbed anticancer drugs were injected preoperatively into the present study, the adsorption isotherm of activated carbon adsorbed aclarubicin (ACR) was measured and its stability in solution was investigated. The adsorption isotherms of 5-FU and MMC were prepared and compared with that of aclarubicin. In the adsorption isotherm of CH-40 activated carbon, the amount of ACR adsorbed was better than that of 5-FU and MMC. The amount of ACR adsorbed for CH-1500 activated carbon was also better than that for CH-40 activated carbon. The stability of ACR adsorbed on activated carbon solution decreased as the concentration of the activated carbon increased. The stability when the activated carbon and ACR concentration ratio was constant in saline solution was 100% after 2 hours and 40% after 7 days.