超声内镜探头引导下乙醇瘤内注射治疗胰腺癌的实验研究

目的 探讨超声内镜探头引导下乙醇瘤内注射治疗胰腺癌的疗效.方法 原位胰腺癌裸鼠20只随机表法随机分为2组,实验组行95％乙醇注射治疗,对照组注射生理盐水,比较治疗前及治疗后24 h血清淀粉酶变化.分别于治疗前及治疗后第7天行超声内镜探头测量肿瘤大小,计算相对肿瘤体积和相对肿瘤增殖率.光学显微镜下观察治疗前及治疗1周后肿瘤组织病理改变.结果 与治疗前相比,实验组治疗后24 h血清淀粉酶出现轻度增高[(2873.5±958.9)U/L比(2004.3±402.0)U/L,P＞0.05],但与对照组治疗后24h的[(2066.1 ±396.6) U/L]比较,差异无统计学意义(P=0.061).实验组治疗后第7天相对肿瘤体积明显低于对照组[(0.45±0.08)比(1.25 ±0.06),P＜0.01)],相对肿瘤增殖率为36％.治疗1周后实验组病理组织学显示肿瘤坏死变性严重,有大面积凝固性坏死;而对照组仅见少量坏死.结论 95％乙醇瘤内注射可造成肿瘤坏死,在裸鼠原位胰腺癌模型中有一定的治疗效果.     

离体猪胰腺钬激光消融的实验研究

目的 探讨不同能量、频率、时间的钬激光对离体猪胰腺的消融范围,建立钬激光对离体猪胰腺消融的回归方程,为临床内镜超声引导下钬激光对胰腺肿瘤的消融治疗提供实验基础.方法 根据预实验确定钬激光能量、时间、频率3个参数可取值的范围,每个参数分别取5个数值,设计一个包含125种不同组合的随机化表,每个组合参数进行一次离体猪胰腺的消融实验,共125次.应用内镜超声探头观察消融灶的声像图,测量消融范围,切下消融部位组织行病理检查.结果 超声见消融部位呈云雾状高回声区.肉眼观消融灶截面近似椭圆球体,中间为碳化,外层为灰白色坏死区域.镜下见消融灶中间为“水池样”空腔,周围为胰腺组织凝固性坏死区,外周有炎症细胞浸润,边缘为正常的胰腺组织.钬激光的作用时间、频率、能量均呈正方向影响消融范围,其中频率影响最大,其次是时间,再次是能量,且差异均具有统计学意义（P值均＜0.01）,最优的参数组合是25 s,25 Hz,1.8J.建立一个包含能量、时间及频率的回归方程,ln（消融体积）=β0＋β1×时间＋β2×频率＋β3×能量（β为回归系数）.结论 钬激光作用于离体猪胰腺可产生明显的组织坏死,所建立的钬激光消融回归方程可指导临床实践.     

内镜超声引导下胰腺射频消融治疗最佳功率的实验研究

目的探索内镜超声引导下猪胰腺射频消融治疗的最佳功率。方法6只健康活体猪按随机数字表法随机分为2组,分别采用内镜超声引导下经胃壁穿刺（超声内镜组）和开腹直视（开腹直视组）的方式对胰腺进行射频消融,各组预设功率分别为5w、10w和15w,监测射频消融过程中阻抗变化情况,射频消融结束时记录消融时间,采用超声内镜和肉眼直视测量胰腺凝固性坏死灶宽径,并对胰腺组织行病理观察。结果6只猪均顺利完成射频消融,术中猪生命体征平稳,无并发症征象。病理均显示胰腺凝固性坏死灶形成,相同射频消融功率下超声内镜组和开腹直视组胰腺凝固性坏死灶宽径肉眼测量值接近,均在10w功率下最大,分别为11．0mm和10．0mm。结论内镜超声引导下猪胰腺射频消融的最佳功率为10w,在此基础上可进一步探索临床胰腺射频消融的最佳功率。     

原位注射法建立人胰腺癌SW1990裸鼠种植瘤模型及高频内镜超声探头对其的监测

目的 建立裸小鼠人胰腺癌原位种植瘤模型,探索监测种植瘤生长的方法.方法 将对数生长期的人胰腺癌细胞株SW1990制备成细胞悬液,原位注射于Balb/c-nu裸小鼠胰腺尾部包膜下,利用高频内镜超声(EUS)探头体表观察肿瘤结节的生长及声像图像.结果 20只裸小鼠均接种成功,1只裸鼠于接种后25 d时死亡.接种后14 d,EUS检查的瘤体大小为(8.09±2.61)mm3,肿瘤结节呈均质低回声,边界清楚,周边有包膜及声晕,形态规则,30%的肿瘤结节周边可见低速环绕彩色血流信号;接种后28 d,瘤体增大至(12.40±3.51)mm3,70%的肿瘤结节呈不规则形,部分为分叶状,肿块呈低回声,不均质,未见液化坏死区,70%的肿瘤结节周边可见低速环绕彩色血流信号.结论 原位注射法是建立裸小鼠人胰腺癌原位种植瘤模型较理想的方法,操作简便,成瘤率高;高频内镜超声显像是可靠的监测胰腺原位种植瘤的手段.     

胰腺实性病变的超声内镜消融治疗

胰腺实性病变可分为良性和恶性。近年来，随着影像技术的发展和超声内镜（endoscopic ultrasound，EUS）的普 及，其发病率呈上升趋势。EUS引导下的消融治疗作为一种微创技术，已显示出部分替代手术治疗胰腺实性病变的 潜力，如胰岛素瘤、无功能性胰腺神经内分泌肿瘤和局部进展期的胰腺癌。文章将从EUS引导下消融治疗胰腺神经 内分泌肿瘤及胰腺癌作一简述。     

术中超声引导下射频消融治疗晚期胰体尾癌的临床研究

目的探讨术中超声引导下射频消融治疗晚期胰体尾癌的临床意义。方法对14例手术不能切除的晚期胰体尾癌行术中超声引导下射频消融治疗,观察术中、术后并发症、近期疗效及生存情况。结果所有病例术中均在超声引导下避开大血管进行消融治疗,在超声图中见到胰腺组织明显气化,肉眼观察到胰腺组织明显碳化,病理活检为凝固性坏死。所有病例均无出血、感染、胰瘘等并发症发生,术后癌性腹痛均立即明显缓解,生存期内癌性腹痛完全消失7例,尚能忍受5例,不能忍受2例。平均随访18个月,5例死亡,9例带瘤生存。结论术中超声引导下射频消融治疗晚期胰体尾癌是一种安全有效的姑息性疗法。     

术中超声引导下射频消融治疗晚期胰体尾癌的临床研究

目的 探讨术中超声引导下射频消融治疗晚期胰体尾癌的临床意义.方法 对14例手术不能切除的晚期胰体尾癌行术中超声引导下射频消融治疗,观察术中、术后并发症、近期疗效及生存情况.结果 所有病例术中均在超声引导下避开大血管进行消融治疗,在超声图中见到胰腺组织明显气化,肉眼观察到胰腺组织明显碳化,病理活检为凝固性坏死.所有病例均无出血、感染、胰瘘等并发症发生,术后癌性腹痛均立即明显缓解,生存期内癌性腹痛完全消失7例,尚能忍受5例,不能忍受2例.平均随访18个月,5例死亡,9例带瘤生存.结论 术中超声引导下射频消融治疗晚期胰体尾癌是一种安全有效的姑息性疗法.     

胰腺囊性病变的内镜诊治

随着影像学技术的进展,胰腺囊性病变诊断率逐年增高。胰腺囊性病变是一组异质性的疾病,主要包括导管内乳头状黏液肿瘤、黏液性囊性肿瘤、浆液性囊性肿瘤和其他罕见的囊性病变,不同类型的胰腺囊性肿瘤具有不同的生物学行为,恶变率也完全不同。对胰腺囊性肿瘤患者进行精准诊断,进而决定部分患者行手术治疗,而另一部分患者可行随访观察。超声内镜和超声内镜引导下细针穿刺囊液分析对胰腺囊性疾病的诊断和鉴别诊断发挥着重要作用。超声内镜引导下的胰腺囊性疾病消融治疗,虽然已经开展了十多年,仍处于起步阶段。但该手术安全有效,可作为外科手术的替代疗法,是一项颇具前景的治疗手段。     

超声内镜引导下微波消融猪肝脏、胰腺的试验研究

【摘要】 目的 通过对猪肝脏、胰腺行超声内镜引导下微波消融（EUS guided microwave ablation,EUS-MWA）术后的机体生理状态及局部病理改变的观察,探讨其可行性与安全性。 方法 健康小型猪8头。全麻后插入超声内镜,在胃内扫查定位肝脏、胰腺,经超声内镜钳道插入直径1.9mm微波消融针消融肝脏和胰腺。肝脏消融功率设置为65W,时间10min;胰腺消融功率设置为60W,时间5min。术前及术后检测肝转氨酶、血清淀粉酶生化指标,并于术后当天行腹部CT平扫观测消融范围及并发症情况。术后6h、24h各处死2头观察穿刺路径上有无重要结构损伤。其余4头饲养至术后2周,观察术后动物饮食、活动、精神状态。对解剖后肝脏、胰腺组织消融区进行病理分析,以未消融区组织做对照。 结果 8头小型猪均顺利接受超声内镜引导下肝脏、胰腺微波消融术,消融灶18处（肝脏10处,胰腺8处）,除一例胰腺定位困难,其他手术过程顺利,术中动物生命体征平稳。术后当天CT平扫提示肝脏及胰腺出现类圆形低密度灶（肝脏最大径2.8±0.3cm,胰腺最大径2.0±0.2cm）,未见腹腔游离气体及胸腹腔积液。术后6h血淀粉酶开始升高,术后 12-24ｈ出现峰值,肝酶仅轻度升高。术后继续饲养组动物出现短时间拒食及活动减少,但均于术后1-2天开始逐渐恢复正常,未见发热、呕吐等异常表现。处死组动物除一例胰腺消融时发生胃壁灼伤外,其他均无穿刺道灼伤、邻近器官损伤及出血情况。病理观察可见肝脏消融中心大片凝固性坏死及周围出血区;胰腺消融区组织可见散在灶状坏死。 结论 本试验初步证实EUS-MWA消融猪肝脏、胰腺安全可行,未来有望用于人体肝脏、胰腺疾病的治疗。     

EUS引导下介入治疗现状

内镜超声检查术(endoscopic ultrasonography,EUS)引导下介入治疗是目前EUS发展的重要方向,主要包括EUS引导下腹腔神经丛阻滞术(EUS-guided celiac plexus neurolysis,EUS-CPN)、EUS引导下胰腺假性囊肿引流、EUS引导下胆管引流(EUS guided biliary drainage,EUS-BD)、超声内镜引导下肿瘤注射治疗等.EUS-CPN效果优于传统镇痛治疗;无论是首次CPN治疗是否有效,再次进行CPN治疗价值有限;选择单侧注射还是双侧注射与操作者习惯及熟练程度有关,在治疗效果上目前没有明显差异.EUS引导下假性囊肿引流为符合引流条件的患者首选方法.EUS-BD的有效性和安全性基本已经得到了认可.EUS引导下酒精注射治疗目前是不适合手术或不愿意接受手术的胰腺神经内分泌肿瘤患者最佳的选择;内镜超声引导下射频消融与EUS引导下胰腺癌注射治疗价值有待进一步证实.     

^125I粒子组织间植入对人胰腺癌裸鼠移植瘤PCNA、VEGF表达的影响

背景：组织间近距离放射疗法是一种有效局部控制胰腺癌的方法,目前已用于前列腺癌、乳腺癌、脑胶质瘤、舌癌、直肠癌等的临床治疗。目的：观察^125I粒子组织间植入对人胰腺癌裸鼠移植瘤增殖细胞核抗原（PCNA）和血管内皮生长因子（VEGF）表达的影响,探讨其治疗胰腺癌的分子机制。方法：BALB／c裸鼠腋下接种人胰腺癌SW1990细胞。按不同剂量率将裸鼠分为2．18cGy／h组、4．46cGy／h组、5．62oCy／h组和7．26cGy／h组,通过TPS计算各治疗组所需植入粒子的放射活度和数量,并设立相应对照组。粒子植入后每4d测量肿瘤体积．21d后处死所有裸鼠,取胰腺癌移植瘤组织行HE染色,以免疫组化SP法检测移植瘤组织PCNA和VEGF表达。结果：治疗组肿瘤体积增长缓慢,可见大片坏死;而对照组肿瘤体积增长迅速,无明显坏死。2．18cCy／h组、4．46cGy／h组、5．62cGy／h组和7．26cGy／h组PCNA（49．7％±6．0％、37．8％±3．6％、30．6％±2．1％和20．8％±2．3％）和VEGF（17．1％±2．7％、9．4％±1．5％、7．4％±0．5％和3．6％±0．9％）阳性率逐渐减弱,且均显著低于相应对照组（P〈0．01）。结论：^125I粒子组织间植入人胰腺癌裸鼠移植瘤可引起肿瘤组织坏死,明显抑制肿瘤生长,可能与其诱导肿瘤组织PCNA和VEGF表达降低有关。     

发光二极管光动力学疗法对裸鼠肺移植瘤的抑制作用研究

目的 探讨以发光二极管（LED）作为光动力学疗法（PDT）光源对裸鼠肺移植瘤的抑制作用.方法 将15只雄性裸鼠随机分为荷瘤对照组、单纯激光照射组和LED-PDT治疗组,各5只.建立肺腺癌A549裸鼠移植瘤模型,荷瘤对照组不予任何治疗;单纯激光照射组不予光敏剂;LED-PDT治疗组腹腔内注射photosan-3,然后进行肿瘤局部PDT,PDT后裸鼠仍避光7 d.测量治疗前后裸鼠肿瘤体积和组织学形态变化.结果 治疗后,荷瘤对照组和单纯激光照射组肿瘤体积均大于LED-PDT治疗组（P＜0.05）.LED-PDT治疗组镜下病理组织切片见细胞坏死、崩解、细胞轮廓模糊;其他两组肿瘤表面无明显变化,病理显示肿瘤组织无坏死.结论 LED-PDT可以使荷瘤裸鼠肿瘤生长速度明显变慢,肿瘤组织局部切片可见肿瘤细胞坏死,PDT可用于抑制活体肺腺癌的生长速度.     

EUS-Guided Antitumor Therapy for Pancreatic Tumors

Endoscopic ultrasound (EUS) is a very useful modality for the diagnosis and staging of pancreatic masses. With the advent of EUS-guided fine-needle aspiration technology, this modality has made a tremendous leap from imaging modality to histologic diagnosis and therapeutic intervention. EUS offers high-resolution images of and unparalleled access to the pancreas. After locating the tip of the echoendoscope in the duodenum or stomach, several drugs or local treatment modalities can be delivered directly into the pancreas. EUS-guided ethanol lavage with/without paclitaxel injection has been tested for the treatment of cystic tumors of the pancreas, with complete resolution of cystic tumor being observed in up to 70-80% of patients. Ethanol injection is also performed for the management of solid neuroendocrine tumors of the pancreas. Various type of EUS-guided injection have also been investigated for the treatment of pancreatic cancer. An activated allogenic mixed lymphocyte culture (Cytoimplant) was injected in patients with advanced pancreatic cancer. A replication-deficient adenovirus vector carrying the tumor necrosis factor-alpha gene was also delivered intratumorally by EUS. ONYX-015 is an oncolytic attenuated adenovirus that exhibits replication preferentially in malignant cells, causing cell death, and this has also been injected into pancreatic cancers under EUS guidance. EUS-guided local ablation therapies such as radiofrequency ablation, photodynamic therapy, and brachytherapy are also under investigation. EUS-guided fine-needle injection for various solid or cystic lesions is a rapidly expanding field. This article reviews the various applications of EUS for the treatment of pancreatic tumors.     

Pathophysiology of chronic pancreatitis induced by dibutyltin dichloride joint ethanol in mice

AIM: To search for a new chronic pancreatitis model in mice suitable for investigating the pathophysiological processes leading to pancreatic fibrosis.METHODS: The mice were randomly divided into 2 groups(n = 50), control group and model group. The mice in model group were given ethanol(10%) in drinking water after injection of dibutyltin dichloride(DBTC)(8 mg/kg BW) in tail vein. The mice in control group were injected with only solvent into tail vein( 60 % ethanol, 20% glycerine and 20% normal saline) and drank common water. At days 1, 7, 14, 28, and 56 after application of DBTC or solvent, 10 mice in one group were killed at each time point respectively. Blood was obtained by inferior vena cava puncture. The activity of amylase, concentration of bilirubin and hyaluronic acid in serum were assayed. The pancreas was taken to observe the pancreatic morphology by HE staining, and to characterize the pancreatic fibrosis by Masson staining. The expression of F4/80, CD3 and fibronectin(FN) were assayed by immuno-histochemistry or Immunofluorescence technique. Collagen typeⅠ(COL1A1) in pancreas were detected by Western blot. The expression of matrix metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinases-1(TIMP-1) m RNA in the pancreas was assessed by real time PCR.RESULTS: DBTC induced an acute edematous pancreatitis within 1 d. The dilated acini, scattered acinar cell necrosis, and inflammatory cells were found at day 7. Extensive infiltration with inflammatory cells following deposition of connective tissue was observed at day 14. At day 28, level of pancreatic fibrosis was aggravated. The pancreatic tissue was replaced by an extended interstitial fibrosis at the end of 2 mo. There was significant difference in the level of amylase, bilirubin and hyaluronic acid in serum between control group and model group(P 0.05). The level of COL1A1 and FN in pancreas increased. The expression of MMP-1 m RNA in pancreas decreased, but TIMP-1 m RNA increased at model group.CONCLUSION: DBTC joint Ethanol drinking can induce chronic pancreatitis in accordance with the pathophysiological modification of human. DBTC joint Ethanol-induced pancreatitis in mice is an effective and handy experimental method. The model is suitable to study the mechanism of pancreatic fibrosis in chronic pancreatitis.     

Synergetic anticancer effect of combined gemcitabine and photodynamic therapy on pancreatic cancer in vivo

AIM： To investigate the anti-tumor effects of combined cytotoxic drug （gemcitabine） and photodynamic therapy （PDT） on human pancreatic cancer xenograft in nude mice.
METHODS： Human pancreatic cancer cell line SW1990 was used in the investigation of the in vivo effect of combined gemcitabine and PDT on human pancreatic cancer xenograft in mice. Sixty mice were randomly allocated into a control group （without treatment）, photosensitizer treatment group （2 mg/kg photosan, without illumination）, chemotherapy group （50 mg/kg gemcitabine i.p.）, PDT group （2 mg/kg photosan ＋ laser irradiation） and combined treatment group （photosan ＋ chemotherapy）, with 12 mice in each group. Tumor size was measured twice every week. Anti-tumor activity in different groups was evaluated by tumor growth inhibition （TGI）
RESULTS： No significant anti-tumor effect was observed either in photosensitizer treatment group or in chemotherapy group. PDT led to necrosis in cancer lesions and significantly reduced tumor volume compared with photosensitizer on day 6 and at the following time points after initialization of therapy （0.24 ± 0.15-0.49 ± 0.08 vs 0.43 ± 0.18-1.25± 0.09, P 〈 0.05）. PDT significantly reduced tumor volume in combined treatment group compared with photosensitizer treatment group （0.12 ± 0.07-0.28 ± 0.22 vs 0.39 ± 0.15-2.20 ± 0.12, P 〈 0.05）, small dose chemotherapy group （0.12 ± 0.07-0.28 ± 0.12 vs 0.32 ± 0.14-1.16 ± 0.08, P 〈 0.05） and control group （0.12 ± 0.07-0.28 ± 0.12 vs 0.43 ± 0.18-1.25 ± 0.09, P 〈 0.05）. TGI was higher in the combined treatment group （82.42%） than in the PDT group （58.18%）.
CONCLUSION： PDT has a significant anti-tumor effect, which is maintained for a short time and can be significantly enhanced by small doses of gemcitabine.     

塞来昔布抑制肺癌增殖及血管生成的实验研究

目的探讨选择性环氧化酶-2(COX-2)抑制剂塞来昔布(西乐葆)对A549肺癌移植瘤的抑制作用及其对肿瘤组织血管生成的影响。方法建立裸鼠肺癌细胞株A549移植瘤模型,药物组给予含1 250 ppm塞来昔布的饮水,连续28 d,计算抑瘤率。采用免疫组化法研究移植瘤血管内皮生长因子(VEGF),微血管密度(MVD)的表达.。结果药物组平均瘤重(0.65±0.20)g,对照组平均瘤重(1.45±0.30)g,两组有显著性差异(P<0.01),塞来昔布抑瘤率为57.23%。药物组与对照组VEGF积分分别为(0.184±0.022)和(0.295±0.032)(P<0.01),MVD平均值分别为(18.80±3.67)和(34.50±5.20)(P<0.01)。结论塞来昔布对A549肺癌移植瘤有明显抑制作用,其作用机制之一与抑制肿瘤血管生成相关。     

EUS-guided photodynamic therapy of the pancreas: a pilot study

BACKGROUND: Photodynamic therapy of pancreatic cancer by using percutaneously placed light catheters has been reported. The feasibility and safety of EUS-guided photodynamic therapy of the pancreas was studied in a porcine model. METHODS: After injection of porfimer sodium, a 19-gauge needle was inserted into the pancreas, the liver, the spleen, and the kidney under EUS guidance. A small diameter quartz optical fiber was passed through the EUS needle and used to illuminate the tissue with laser light. The tissue response to photodynamic therapy was examined. RESULTS: Localized tissue necrosis was achieved in all organs, without significant complication. There was no significant difference in inflammation induced by photodynamic therapy within the various organs. CONCLUSIONS: EUS-guided photodynamic therapy is a safe and simple technique that can induce small areas of focal tissue ablation within the liver, the pancreas, the kidney, and the spleen, and potentially could be used to treat a variety of benign and malignant conditions.     

量子点-RGD探针光动力疗法联合吉西他滨治疗胰腺癌荷瘤裸鼠初探

背景:胰腺癌发病隐匿,进展迅速,预后极差。光动力疗法(PDT)是20世纪80年代发展起来的一种新型抗肿瘤治疗手段。目前关于PDT在体内靶向治疗胰腺癌的研究甚少。目的:探讨以量子点-RGD(QDs-RGD)探针为光敏剂的PDT联合吉西他滨对胰腺癌移植瘤裸鼠的治疗效应。方法:合成QDs-RGD探针,制备胰腺癌移植瘤裸鼠模型。采用小动物活体成像术观察QDs-RGD、QDs探针注射入模型裸鼠体内1、5、10、24 h后的显影情况。取40只造模成功的裸鼠,随机分为5组:对照组(不给予任何治疗);单纯光照组(激光630 nm,120 J/cm2,持续照射20 min);PDT组(QDs-RGD 0.5 nmol+激光照射);吉西他滨组(吉西他滨50 mg/kg);联合治疗组(QDs-RGD 0.5 nmol+激光照射+吉西他滨50 mg/kg)。第18 d处死全部裸鼠,取出瘤体,称重并测量体积,计算抑瘤率。结果:QDs-RGD注射1 h后,肿瘤显影逐渐清晰,注射5 h后显影达高峰,随后逐渐减弱。QDs于瘤体附近的聚集浓度明显低于QDs-RGD,注射10 h后肿瘤部位已无显影。PDT组、吉西他滨组和联合治疗组的瘤重、瘤体积均显著低于对照组和单纯光照组(P0.01),其中联合治疗组又显著低于PDT组和吉西他滨组(P0.05);对照组与单纯光照组间、PDT组与吉西他滨组间瘤重、瘤体积差异均无统计学意义(P0.05)。联合治疗组、吉西他滨组、PDT组的抑瘤率分别为70.5%、43.5%、37.1%。结论:以QDs-RGD探针为光敏剂的PDT联合吉西他滨能明显抑制裸鼠体内胰腺癌移植瘤的生长,为临床治疗胰腺癌提供了一条新思路。