腹腔镜胃癌手术前后腹腔冲洗液survivin的检测

目的:探讨腹腔镜胃癌根治术对腹腔癌细胞脱落的影响。方法:收集50例胃癌患者的腹腔冲洗液,采用流式细胞术检测胃癌患者腹腔镜手术前后腹腔冲洗液存活素(survivin)的表达情况,并采用薄层液基细胞制片术进行腹腔冲洗液细胞学检查(pleural lavage cytology,PLC)。结果:50例患者腹腔镜胃癌手术前腹腔冲洗液中survivin阳性表达率为58.0%(29/50),术后阳性率为66.0%(33/50),两者差异无统计学意义(P>0.05)。手术前后PLC阳性率分别为22.0%(11/50)和28.0%(14/50)(P>0.05)。术前、术后PLC阳性患者survivin均为阳性。结论:腹腔镜胃癌根治手术未增加腹腔胃癌细胞的脱落。但由于实验样本数量较少,尚需进一步通过长期随访及多中心研究证实。     

腹腔镜胃癌根治术后腹腔冲洗液CEA mRNA的变化和意义

目的探讨腹腔镜胃癌根治术对腹腔脱落癌细胞的影响。方法收集44例行腹腔镜胃癌根治术患者胃癌切除前后腹腔冲洗液，并以RT—PCR方法检测腹腔冲洗液CEAmRNA表达情况。结果腹腔镜胃癌根治术患者CEAmRNA表达胃癌未切除时阳性率为27．3％（12／44），切除后阳性率为36．4％（16／44），两组之间差异无统计学意义（P〉0．05）。结论腹腔镜胃癌根治术不增加腹腔胃癌细胞的脱落。     

结直肠癌患者手术中腹腔及肠腔脱落细胞学研究

目的 结肠直肠癌腔及肠腔脱落细胞的定性研究,为术中无瘤接触、常规腹及远端结直肠冲洗提供理论依据。方法随机选择２３例手术治疗的结直肠癌患者,２２例术中切除肿瘤后,作肠吻合前用５００－１０００ｍｌ生理盐水行远端结直肠冲洗,其中１４例肿瘤侵及浆膜,开腹后用１００－２００ｍｌ生理盐水行腹腔冲洗;１例仅行腹腔冲洗,未行肠腔冲洗。收集冲洗液迅速送检作脱落细胞学检查。结果 行腹腔冲洗的１５例中１例找到癌细胞,阳     

腹腔镜胃癌手术对胃癌细胞腹腔种植转移影响的临床研究

目的探讨腹腔镜胃癌手术和开腹胃癌手术对胃癌细胞种植转移的影响。方法收集2006年4月至2006年11月间腹腔镜胃癌手术中的45例和开腹胃癌手术中的41例患者术前、术后腹腔冲洗液各100ml,进行腹腔脱落肿瘤细胞学检查。腹腔镜组,气腹后通过套管针侧孔引出CO2气体,经过100ml生理盐水的滤过后,收集过滤液进行肿瘤细胞学检查。腹腔镜组术后用100ml生理盐水冲洗腹腔镜手术器械,进行腹腔镜器械冲洗液肿瘤细胞学检查。所有液体离心取沉淀,涂片固定HE染色,光镜下观察肿瘤细胞。结果腹腔镜气腹CO2气体滤过液中未观察到肿瘤细胞;腹腔镜器械冲洗液发现3例阳性(6.7%);两组术前腹腔冲洗液中肿瘤细胞的阳性率分别为60.0%和60.9%,术后肿瘤细胞阳性率分别为53.3%和56.1%,两种手术方式术前、术后腹腔冲洗液肿瘤细胞无明显差异。腹腔冲洗液与胃癌TNM分期有关,两组肿瘤细胞Ⅲ期阳性率均显著高于Ⅱ期,具有非常显著统计学差异(P0.01),即两组中胃癌细胞阳性检出率在肿瘤的病理分期越晚,阳性率越高。结论 CO2气腹不会引起肿瘤细胞的播散。腹腔镜术中器械污染是引起切口转移的主要原因之一。腹腔镜手术不增加肿瘤细胞播散种植的机会。     

应用分支DNA技术及SqRT-PCR方法检测结直肠癌患者腹腔冲洗液中游离癌细胞

目的探讨分支DNA(b-DNA)及半定量RT-PCR(SqRT-PCR)在结直肠癌术中腹腔冲洗液中游离癌细胞检测中的应用。方法分别采用基于b-DNA信号放大的基因表达定量检测技术及SqRT-PCR方法检测48例结直肠癌患者术中腹腔冲洗液中CEA mRNA的表达,同时行腹腔冲洗液细胞学检查(peritoneal lavage cytolo-gy,PLC),收集12例结直肠良性病变患者的腹腔冲洗液为阴性对照,GAPDH mRNA为内参对照。结果b-DNA技术和SqRT-PCR方法检测游离癌细胞的阳性率(43.8%,31.3%)较PLC的检出率(4.2%)高(P0.01)。结直肠癌患者腹腔冲洗液中CEA mRNA的相对表达量均与肿瘤分化程度、浆膜侵犯程度和Dukes分期有关(P0.05),而与肿瘤大小、患者性别、年龄无关(P0.05)。结论b-DNA技术和SqRT-PCR方法检测游离癌细胞各有优缺点;腹腔内游离癌细胞的存在与结直肠癌临床病理因素有关。     

胃癌腹腔微转移的研究及其临床意义

目的运用细胞学及RT-PCR方法检测胃癌患者术中腹腔冲洗液和腹膜组织,以探讨这些方法对预测胃癌腹腔微转移的意义.方法收集3 8例胃癌和5例胃良性病变患者的术中腹腔冲洗液,并同时切除少量大网膜、腹膜作为对照.用RT-PCR方法测定冲洗液中游离细胞的CEAmRNA表达,同时作冲洗液细胞学检查（PLC）.结果腹腔冲洗液和腹膜组织中的CEA mRNA的阳性率分别为36.8%（14/38）和39.5%（15/38）,皆高于腹腔冲洗液PLC的26.3%（10/38）.CEA mRNA的阳性率与肿瘤的分化程度、浸润深度、淋巴结转移、浆膜侵犯深度有关.结论CEART-PCR方法对于检测腹腔微量游离癌细胞较PLC有更高的灵敏度和特异性,是一种检测胃癌腹腔微转移的有效方法.     

结直肠癌患者手术前后腹腔脱落肿瘤细胞检测的临床意义

目的探讨结直肠癌患者手术前后腹腔脱落肿瘤细胞检出阳性率及其与临床病理分期的关系和对预后的影响。方法对121例结直肠癌患者在手术进入腹腔后和关闭腹腔前,进行腹腔脱落细胞学检查,并与患者的临床病理分期和随访结果进行对比分析。结果全组患者腹腔冲洗液中肿瘤细胞检出阳性者术前为35例(28.9%),术后4例复发(11.4%);术后检出阳性者为13例(10.7%),术后7例复发(53.8%);手术前后均为阴性者其术后复发为2例(2.4%)。病理分期越晚肿瘤细胞检出阳性率越高(均P<0.05或0.00)。结论对结直肠癌患者手术前后进行腹腔冲洗液细胞学检查方法简单,具有一定的临床价值。     

腹腔镜胃癌根治术后腹腔冲洗液CEA mRNA的变化和意义

目的探讨腹腔镜胃癌根治术对腹腔脱落癌细胞的影响。方法收集44例行腹腔镜胃癌根治术患者胃癌切除前后腹腔冲洗液,并以RT-PCR方法检测腹腔冲洗液CEA mRNA表达情况。结果腹腔镜胃癌根治术患者CEA mRNA表达胃癌未切除时阳性率为27.3%(12/44),切除后阳性率为36.4%(16/44),两组之间差异无统计学意义(P>0.05)。结论腹腔镜胃癌根治术不增加腹腔胃癌细胞的脱落。     

直肠癌术中腹腔冲洗液脱落细胞学临床研究进展

直肠癌术后局部复发、盆腔种植转移成为直肠癌术后主要的致死原因。国内外报道直肠癌根治术后复发率为10％～30％，为预防直肠癌术后复发、转移，国内外学者提出了许多有效的方法如全直肠系膜切除（TME），新辅助化疗，术中腹腔冲洗等。其中术中腹腔冲洗作为手术无瘤技术的重要措施已在临床广泛应用并取得明显的成效。在进行腹腔冲洗达到减瘤的目的的同时，术中腹腔冲洗液脱落细胞学的研究渐成热点，现将国内外有关直肠癌术中腹腔冲洗液脱落细胞学的研究综述如下。     

腹腔液胃癌细胞学与CEA mRNA检测比较的意义

目的：探讨胃癌病人术中腹腔冲洗液细胞学及腹腔冲洗液、腹膜组织中的癌胚抗原（CEA）mRNA，以探讨对腹腔中游离癌细胞和预测腹膜转移的诊断价值。方法：收集48例胃癌和5例胃良性病变病人的腹腔冲洗液或腹水，并同时切除少量大网膜、膈腹膜和盆腔腹膜作为对照。冲洗液行常规细胞学检查，并逆转录－聚合酶链反应（RT－PCR）法检测腹腔游离癌细胞和网膜组织中CEA mRNA的表达。结果：腹腔冲洗液和腹膜组织中的CEA mRNA的阳性率分别为39．58％（19／48）和43．75％（21／48），皆高于腹腔冲洗液细胞学27．08％（13／48）（P＜0．05）。CEA mRNA的表达与淋巴结转移、肿瘤浸润深度、浆膜侵犯程度及TNM分期呈正相关。结论：腹腔冲洗液CEA mRNA和细胞学检查是检测腹腔游离癌细胞和预测腹膜转移的有效方法；如同时行腹膜组织CEA mRNA的检测可能更有助于诊断。     

Intraoperative lavage for cytological examination in 1,297 patients with gastric carcinoma.

BACKGROUND: This study examined the clinical value of intraoperative peritoneal lavage for cytological examination in patients with gastric cancer. Peritoneal dissemination is the most frequent mode of recurrence for this tumor. METHODS: A retrospective of lavage findings, other factors, and outcome was performed in 1,297 patients with gastric cancer who underwent intraoperative peritoneal lavage. RESULTS: The 5-year survival rate of patients with positive lavage cytology was only 2%. Patients who underwent curative resection and had negative cytology had a significantly better 5-year survival rate (P < 0.001). Even among patients with macroscopic peritoneal dissemination, the survival rate was significantly better with negative cytology, which reflected fewer free cancer cells in the peritoneal cavity. Serum concentrations of carcinoembryonic antigen and carbohydrate antigen 19-9 were significantly higher in patients with positive cytology. Multivariate analyses indicated that intraoperative cytological findings was an independent prognostic factor for survival, and was the most important factor for predicting peritoneal recurrence. CONCLUSIONS: Intraoperative peritoneal lavage cytology is important in predicting survival and peritoneal recurrence in gastric cancer.     

腹腔镜下进展期胃癌根治术中腹腔游离癌细胞检测的意义

目的探讨腹腔镜下进展期胃癌根治术对腹腔脱落肿瘤细胞检测的影响,并探讨提高腹腔游离癌细胞检出率的方法。方法选取华中科技大学同济医学院附属协和医院胃肠外科35例初治的进展期胃癌病人,行腹腔镜下胃癌根治术。所有病人分别在术中探查前及关腹前各行一次腹腔冲洗,并收集灌洗液。比较两组灌洗液中肿瘤脱落细胞阳性检出率及肿瘤标志物上皮膜抗原(epithelial membrane antigen,EMA)、癌抗原(carbohydrate antigen,CA)125、CA19-9和癌胚抗原(carcinoembryonic antigen,CEA)的表达情况。结果35例进展期胃癌病人探查前腹水或腹腔灌洗液中涂片细胞学检查中检出阳性率为5.7%(2/35),关腹前为5.7%(2/35),两者统计比较差异无统计学意义(P>0.05)。免疫组织化学法检测探查前与关腹前腹水或腹腔灌洗液中EMA的表达(4/35,3/35)比较,差异无统计学意义(P>0.05);两个时间点CA125的表达(1/35,3/35)比较,差异无统计学意义(P>0.05)。探查前腹水或腹腔灌洗液中,腹腔灌洗液脱落细胞学法游离癌细胞检出阳性率为5.7%(2/35),而免疫组织化学法检测EMA的阳性率为11.4%(4/35),两者比较差异有统计学意义(P=0.010);免疫组织化学法检测CA125的阳性率为8.6%(3/35),与腹腔灌洗液脱落细胞学法(2/35)比较,差异有统计学意义(P=0.005);免疫组织化学法联合检测EMA及CA125的阳性率为11.4%(4/35),与腹腔灌洗液脱落细胞学法(2/35)比较,差异有统计学意义(P=0.010)。结论腹腔镜下进展期胃癌根治术并不会增加癌细胞脱落风险,且肿瘤脱落细胞发生率低。但腹腔镜下胃癌根治术后腹腔游离癌细胞的诊断,需要更为有效的检测方法。     

Long-Term Survival in Advanced Small Cell Carcinoma of the Colorectum: Report of a Case

A 47-year-old man was referred to our institution with bloody diarrhea. An endoscopic examination showed a 6-cm tumor with central ulceration in the upper rectum. A high anterior resection of the rectum with lymphadenectomy was performed with the diagnosis of colorectal cancer. An intraoperative cytological examination found many free cancer cells in the peritoneal lavage fluid. Histologically, the tumor had invaded deeply and exposed the serosa, and was diagnosed as a small cell carcinoma of the colorectum with marginal lymph node metastasis. The patient is alive without any evidence of recurrence approximately 37 months after surgery despite the aggressive clinical behavior and a high mortality rate associated with this tumor. This case of an advanced small cell carcinoma of the colorectum showed a good outcome even though the cytology of the peritoneal lavage was positive. Received: January 5, 2001 / Accepted: July 17, 2001     

Prognostic value of peritoneal lavage cytology in gastric cancer

The microscopic detection of free peritoneal tumour cells in peritoneal lavage fluid in gastric cancer patients is a useful predictor of peritoneal recurrence and poor prognosis. The aim of this study was to verify the prognostic significance of intraoperative peritoneal lavage cytology and its value as a predictor of peritoneal recurrence. We evaluated the presence of free peritoneal tumour cells with extemporary cytological examination in a series of 170 peritoneal washing samples from patients undergoing gastrectomy for gastric cancer over the period from January 1992 to June 2001. Twenty-eight patients (16%) had positive extemporary lavage cytology and there were no false-negatives as compared with the final examination. All patients with positive cytology presented serosal infiltration (T3/T4). Positive peritoneal lavage cytology was a predictor of poor prognosis and peritoneal recurrence: the 24 month survival rate was 17% for positive and 60% for negative cases (P = 0.003); in positive cases 71% of recurrences were located in the peritoneum. Intraoperative cytological examination of peritoneal washings can detect the presence of free malignant cells in the peritoneal cavity and can be used to select patients who may benefit from intraperitoneal chemotherapy.     

检测腹腔灌洗液中细胞内端粒酶活性对结直肠癌患者的诊断价值

目的探讨测定腹腔灌洗液中细胞内端粒酶活性对检测腹腔中游离癌细胞的敏感性。方法采用原位分子杂交(ISH)技术测定28例结直肠癌患者术始、术毕腹腔灌洗液中细胞内端粒酶活性,同时与常规细胞学检查结果进行比较。结果28例结直肠癌患者术始、术毕腹腔灌洗液中细胞内端粒酶阳性率分别为42.9%(12/28)和21.4%(6/28),P>0.05;差异无显著性意义。常规细胞学检查游离癌细胞阳性率术始、术毕分别为14.3%(4/28)和3.6%(1/28);两者差异也无显著性意义(P>0.05)。术始,腹腔灌洗液中细胞内端粒酶阳性率为42.9%,细胞学检查阳性率为14.3%,两者差异有显著性意义(P<0.05);术毕,腹腔灌洗液中细胞内端粒酶阳性率为21.4%,细胞学检查阳性率为3.6%,两者差异有显著性意义(P<0.05)。原发癌肿已侵出浆膜层者和原发癌肿侵及和未侵及浆膜层者,术始腹腔灌洗液中细胞内端粒酶阳性率分别为73.3%和7.7%,两者差异有非常显著性意义(P<0.01);腹腔灌洗液中细胞内端粒酶阳性与临床原发癌肿侵出浆膜层呈正相关,但与原发癌肿大体类型、组织学类型及有无淋巴结转移均无关(均P>0.05)。结论用ISH技术测定腹腔灌洗液中细胞内端粒酶活性对检测腹腔中游离癌细胞的敏感性明显高于常规细胞学检查。     

Intraoperative Pleural Lavage in Esophageal Carcinoma

Background: Cytological examination of intraoperative pleural or peritoneal lavage specimens is useful for predicting outcomes for patients with various carcinomas. There have been few reports regarding cytological examination of pleural lavage fluid in esophageal carcinoma.Methods: Intraoperative pleural lavage fluid was collected before and after esophagectomy and was examined by Papanicolaou and Giemsa staining for 78 patients with esophageal carcinoma.Results: Although epithelial cells were found for 29 patients, only blood cells were detected for 48. The remaining one patient exhibited no cells in the specimen. For 4 of 78 (5.2%) patients, tumor cells were detected in the pleural lavage fluid after esophagectomy. Three of these four patients had T4 tumors.Conclusions: Positive cytological findings for pleural lavage fluid, using Papanicolaou and Giemsa staining, is correlated with regrowth of residual tumor and poor prognosis in esophageal carcinoma.     

Effectiveness of intraoperative cytological examination of peritoneal washings for patients with pancreatic cancer]

To confirm the effectiveness of cytological examination of peritoneal washings for detecting invisible micro-peritoneal dissemination in patients with pancreatic cancer, results were analyzed with the survival and the background factors of the patients. Cytological examination of peritoneal washings or ascitic fluid at recto-vesical pouch or pouch of Douglas was performed in 37 patients with primary pancreatic cancer. Positive results for cancer cells were obtained in five of 9 patients (55.6%) who received cytological examination of ascitic fluid and in seven of 28 patients (25.0%) who received that of peritoneal washings. Four of 6 patients (66.7%) with visible peritoneal dissemination showed positive results. These 6 patients died of peritoneal dissemination with about 10 months. Eight of 31 patients (25.8%) without visible peritoneal dissemination showed positive results of the cytological examination. Two of the 8 patients received resection of the tumor. Other 6 patients without resection developed clinically evident peritoneal carcinomatosis. A high positive rate (66.7%) of cytological examination of the patients with visible peritoneal dissemination and a high incidence of appearance of peritoneal carcinomatosis in patients with positive cytological results but without visible peritoneal dissemination (75.0%; positive vs 26.1%; negative) indicate a high reliability of the cytological examination to detect invisible micro-peritoneal dissemination.     

Intraoperative pleural lavage cytology in lung cancer patients

Cytology of intraoperative pleural lavage was examined in 164 lung cancer patients who underwent pulmonary resections. None of the patients had any pleural effusion or dissemination. Cytology was performed three times: (1) at thoracotomy, (2) immediately after resection, and (3) after washing the pleural cavity with 5,000 mL of physiological saline solution just before closure of the chest wall. Twenty-three patients (14%) had more than one positive cytological finding. The frequency of positive cytological findings was significantly correlated with pathological T classification, pleural status, and pathological stage. The positive lavage group had a significantly higher recurrence rate than the negative lavage group in patients with stage I or stage II cancer. Four patients in the positive lavage group (17.4%) had recurrence in pleura or pericardium whereas only 1 patient in the negative lavage group (0.7%) had a recurrence in pericardium. The positive cytological finding of pleural lavage has more important meaning as a prognostic factor in stage I and stage II and indicates a greater possibility of recurrence in pleura or pericardium, but further examinations to evaluate the viability of detected malignant cells are required so that the positive cytological findings of pleural lavage can be regarded as subclinical pleural dissemination.     

Lack of prognostic role of pre- and postoperative peritoneal cytology and cytokeratin PCR-expression on local recurrence after curative anterior resection for mid-low rectal cancer

Local recurrence continues to be a major problem in rectal cancer. After cancer removal, detection of viable cancer cells could be useful to identify patients at risk for local recurrence. Thus, aim of the study was the detection of residual peritoneal cancer cells with a possible prognostic role for local recurrence. Twenty-nine patients were operated (R0) for low (extraperitoneal) rectal cancer, without neoadjuvant radiochemotherapy. Before and immediately after cancer removal, a peritoneal lavage was done to evaluate by RT-PCR the cytokeratin 20 mRNA on isolated cells and in order to detect cancer cells by the Thin-prep test. After a median follow-up of 39 months, 5 patients died (17%), one for non-cancer-related disease, two (7%) for local recurrence and peritoneal carcinosis, and two for distant metastases. Preoperative cytology with Thin-prep test was positive in 4 patients (14%), while postoperative peritoneal cytology was positive only in 1 patient, different from the previous. No patient developed local or distal recurrence and all were disease-free at the end of the follow-up. RT-PCR analysis was positive on the peritoneal lavage after cancer removal in 11 patients. One died for unrelated cause and no one developed local recurrences. Local recurrence occurred in only 1 of the 2 patients with positive RT-PCR analysis on the first lavage and negative on the second lavage. Our study demonstrates a not important prognostic role of Thin-prep test and RT-PCR of cytokeratin 20 mRNA on the detection of patients at risk for local recurrence after curative resection of rectal cancer.     

肠内营养用于术前肠道准备对结直肠癌患者腹腔和肠腔脱落癌细胞及复发转移的影响

目的探讨术前肠内营养代替肠道准备对结直肠癌患者腹腔和肠腔脱落癌细胞及转移复发的影响。方法2007年3月至2011年12月问前瞻性人组120例结直肠癌者,按随机数字表法随机分为试验组和对照组,每组60例。试验组于术前3d起每日口服30ml／kg肠内营养液,不进行灌肠,不口服泻药和抗生素;对照组采用传统肠道准备,包括术前禁食、口服抗生素和清洁灌肠。两组患者均于开腹后用400ml生理盐水冲洗肿瘤所在局部腹腔,抽取腹腔冲洗液200ml,迅速送检;并于肿瘤切除后肠吻合前,用1000ml生理盐水灌洗远端结直肠腔,收集肠腔冲洗液500ml,迅速送检。比较两组患者腹腔和肠腔内脱落的癌细胞阳性率及术后并发症发生率和复发转移率。结果试验组患者腹腔和肠腔内脱落癌细胞阳性率分别为8．3％（5／60）和15．0％（9／60）．明显低于对照组患者的12．5％（13／60）和31．7％（19／60）（P=0．041和P=O．031）。试验组和对照组分别有55例和57例患者完成随访,随访时间16—46个月;两组局部复发率[5．5％（3／55）比7．0％（4／57）,P=O．733]、远处转移率[10．9％（6／55）比10．5％（6／57）,P=O．984]和3年总体生存率（80％比78％,P=O．312）的差异均无统计学意义。结论术前肠内营养可减少结直肠癌患者腹腔和肠腔脱落癌细胞的阳性率,但并不影响术后局部复发率和远处转移率。