p53基因治疗对鼻咽癌患者局部免疫及预后的影响

目的探讨p53基因治疗后鼻咽癌患者的局部免疫变化及其对肿瘤复发和转移的影响。方法63例经病理确诊的初治中、晚期鼻咽癌患者分为p53治疗组[32例,采用p53腺病毒(r Ad-p53)鼻咽部瘤体内注射和同步放、化疗]和常规治疗组(31例,采用同步放、化疗)。采用免疫组织化学方法检测治疗前、后鼻咽部肿瘤浸润淋巴细胞的T细胞受体CD3和B细胞受体CD20的表达情况,了解肿瘤组织的局部免疫状态;统计患者在治疗后定期回院的复查结果以及随访资料,了解患者的生存率及肿瘤的复发和转移情况。结果p53治疗组治疗后肿瘤组织中的CD3表达比治疗前明显增加(P0.05),治疗前、后CD20的表达差异无统计学意义(P0.05)。p53治疗组和常规治疗组复发和转移率差异有统计学意义(P0.05),p53治疗组5年无瘤生存率优于常规治疗组(P0.05)。结论 p53基因治疗后鼻咽部肿瘤中的肿瘤浸润淋巴细胞增殖以T细胞为主,治疗后的局部细胞免疫状态较治疗前有明显提高,肿瘤的复发和转移率降低,基因治疗联合放化疗可能有助于改善鼻咽癌的预后。     

内镜超声引导下瘤内注射重组人p53腺病毒治疗胰腺癌的短期临床观察

目的 探讨重组人p53腺病毒(rAd-p53)瘤内注射治疗胰腺癌的临床可行性,并对相关文献做一回顾复习.方法 2006年7月5日至12月4日期间,共有8例中晚期胰腺癌患者行内镜超声引导下的穿刺,并经穿刺针于肿瘤内注射rAd-p53注射液.每周注射1次,共4 ～ 5周.rAd-p53治疗的同时,行健择静脉化疗.结果 8例患者全部完成内镜超声下胰腺肿瘤内的rAd-p53注射治疗,平均操作时间为10 min.注射后1例出现面部潮红、胃部灼热感,2例出现发热,体温37.5℃,均自行缓解.死亡1例,7例患者治疗后疼痛明显好转,其中1例疼痛消失,2个月体重增加2 kg.影像学检查所有患者肿瘤均稳定,无明显增大.结论 内镜超声引导下rAd-p53肿瘤内注射安全、可靠,值得进一步研究.     

内镜超声引导下瘤内注射重组人p53腺病毒治疗胰腺癌的短期临床观察

目的探讨重组人p53腺病毒(rAd-p53)瘤内注射治疗胰腺癌的临床可行性,并对相关文献做一回顾复习。方法2006年7月5日至12月4日期间,共有8例中晚期胰腺癌患者行内镜超声引导下的穿刺,并经穿刺针于肿瘤内注射rAd-p53注射液。每周注射1次,共4～5周。rAd-p53治疗的同时,行健择静脉化疗。结果8例患者全部完成内镜超声下胰腺肿瘤内的rAd-p53注射治疗,平均操作时间为10min。注射后1例出现面部潮红、胃部灼热感,2例出现发热,体温37.5℃,均自行缓解。死亡1例,7例患者治疗后疼痛明显好转,其中1例疼痛消失,2个月体重增加2kg。影像学检查所有患者肿瘤均稳定,无明显增大。结论内镜超声引导下rAd-p53肿瘤内注射安全、可靠,值得进一步研究。     

瘤内淋巴管生成与喉鳞癌淋巴转移的关系

选择我喉鳞状细胞癌（LSCC）标本60例，行双重免疫组化染色法计数喉鳞癌瘤内淋巴管。发现喉鳞癌瘤内有新生淋巴管，瘤内淋巴管密度（LVD）与N分期及临床分型存在相关关系。认为瘤内LVD的增高与肿瘤淋巴转移有关。     

p53基因联合化疗治疗复发鼻咽癌1例报告

p53肿瘤抑制基因与肿瘤的发生、发展以及肿瘤病人的预后关系密切，已经用于头颈部鳞癌和非小细胞性肺癌基因治疗的临床试验。国内有用重组腺病毒-p53（rAd-p53）基因瘤内注射结合放疗治疗鼻咽癌，疗效显著，但仍未见关于p53基因联合化疗治疗复发性鼻咽癌的报道。我们试用p53基因联合化疗治疗复发性鼻咽癌1例，取得可喜疗效。     

沙培林注射治疗淋巴管瘤52例临床分析

选取门诊及住院淋巴管瘤52例,采用沙培林注射治疗;对所有病例进行随访,并记录其疗效及毒副作用。结果囊状淋巴管瘤40例,治愈30例（75％）,显效6例（15％）;海绵状和混合型淋巴管瘤12例,治愈2例（16．7％）,显效4例（33．3％）。认为沙培林注射治疗囊状淋巴管瘤效果明显,海绵状和混合型淋巴管瘤疗效较差,可用于无法手术切除部位或器官的海绵状和混合型淋巴管瘤。     

p53基因多态性与鼻咽癌遗传易感性的关系

目的研究广西南宁鼻咽癌患者p53基因遗传多态性与鼻咽癌发生的关系。方法采用病例-对照研究方法,以200例鼻咽癌患者、200例对照人群为研究对象。选取p53基因SNP rs117562731位点作为遗传标记,用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)和测序方法检测rs117562731位点基因型频率和等位基因频率,比较两组不同基因型与鼻咽癌易感性的关系。结果通过对rs117562731位点多态基因型检测分型发现,在鼻咽癌组和对照组CC、CT、TT基因型频率分别为80.0%、17.0%、3.0%和93.0%、5.5%、1.5%。rs117562731位点等位基因及基因型频率在鼻咽癌组与对照组间分布有显著性差异(P0.05)。结论 p53基因SNP rs117562731位点多态性与鼻咽癌之间存在显著的相关性。     

巨噬细胞对动脉壁淋巴管生成的调节作用

巨噬细胞在动脉粥样硬化中发挥多重作用。动脉粥样硬化的进展与病变部位动脉的淋巴管的形态和功能改变有关,但其机制尚不完全清楚。文章主要对动脉粥样硬化中巨噬细胞的来源、分型、标志物及对动脉粥样硬化的功能作用,淋巴管的起源、结构功能及标志物,动脉粥样硬化病变不同时期动脉壁淋巴管生成的变化,淋巴管生成在动脉粥样硬化中的功能作用,巨噬细胞的淋巴管迁移及其参与淋巴管新生的作用机制进行综述,以期为动脉粥样硬化的机制研究和临床治疗提供依据。     

肿瘤淋巴管生成的调控

侵袭和转移是恶性肿瘤的基本特征,淋巴管是恶性肿瘤侵袭转移到局部淋巴结或远处器官的主要途径之一.近年来,越来越多的研究表明肿瘤中存在活跃的淋巴管生成,新生的淋巴管对肿瘤的转移起着重要作用.淋巴管的生成是一个由多种因子参与的复杂过程,除趋化因子、黏附分子和细胞外基质等作用外,淋巴管内皮细胞特异性转录因子和受体等发挥着关键性作用.明确淋巴管生成的调控机制对肿瘤的治疗有重要意义.     

明胶海绵对小鼠肌肉组织血管淋巴管生成的影响

目的观察明胶海绵对小鼠肌肉组织血管淋巴管形成的影响。方法用明胶海绵填充小鼠下肢肌肉组织内。植入后第1、2、3、4、5周解剖小鼠,观察海绵中血管淋巴管形成情况。结果海绵表面可见血管覆盖,并有分支深入其中;海绵与肌肉组织相容性良好。切取的海绵透光可见血管网络。第5周海绵基本吸收,体积明显变小。HE染色显示海绵中央与肌肉海绵交界处管状结构数目先增加后减少,而平均管径逐渐变大（P均〈0.05）。结论明胶海绵可促进小鼠肌肉组织血管与淋巴管生成。     

Effect of rAd-p53 on glucose metabolism in diet-induced and genetic T2DM mice

正Objective To explore the possible mechanism of rAd-p53 involved in glucose metabolism by observing the different effects of rAd-p53 on high-fat diet(HFD)fed with low-dose streptozotocin(STZ)treated(HFD/STZ)mice(β-cell dysfunction model)and db/db mice(genetic insulin resistance model).Methods The established HFD/STZ and db/db diabetic mice were respectively and     

重组人p53腺病毒抑制肝癌HepG2细胞增殖机制的研究

目的探讨重组人p53腺病毒(rAd-p53)抑制肝癌HepG2细胞增殖的作用机制.方法以高、中、低浓度(分别为3×1012VP、3×1011VP和3×1010VP)的rAd-p53处理肝癌HepG2细胞(分别为观察Ⅰ、Ⅱ、Ⅲ组),用免疫组化法检测周期素D1(cylinD1)、周期素E(cyclinE)蛋白在肝癌HepG2细胞中的表达,用MTT比色法观察rAd-p53对肝癌HepG2细胞生长的抑制作用,用流式细胞仪检测细胞周期分布;另设空白对照组.结果①rAd-p53对HepG2细胞有明显的抑制效应,且具有时间、剂量依赖关系.②观察Ⅰ、Ⅱ、Ⅲ组G1期细胞分别上升至75.8%、70.1%、66.5%,明显高于对照组(P<0.05);观察Ⅰ、Ⅱ组S期细胞分别下降至13.2%、20%,明显低于对照组(P<0.05).③观察Ⅰ、Ⅱ、Ⅲ组肝癌HepG2细胞cyclinD1蛋白水平分别减少45.6%、35.1%和26.3%,与对照组比较P均<0.05;cyclinE蛋白水平分别减少33.3%、27.1%和18.1%,与对照组比较P均<0.05.结论 rAd-p53对肝癌HepG2细胞有明显的抑制效应,其抑制效应具有时间、剂量依赖关系,其机制可能是通过抑制下游cyclinD1、cyclinE表达阻止细胞进入S期.     

重组人p53腺病毒基因经介入途径治疗兔VX2肝癌的研究

目的 观察重组人p53腺病毒基因(rAd-p53)经介入途径治疗兔VX2肝癌的效果.方法 将30只新西兰大白兔采用移植法建立兔VX2肝癌模型,并随机均分为A、B、C、D、E5组,于第11天分别行磁共振成像、CT检查,计算肿瘤体积后对各组模型处理,A组:经肝动脉灌注等渗盐水0.5ml/只;B组:经肝动脉灌注超液态碘油0.5ml/只;C组:经肝动脉灌注rAd-p531×106/VP;D组:经肝动脉灌注rAd-p53 1×106/VP加超液态碘油0.5 ml/只; E组:肿瘤内直接注射rAd-p53 1×106/VP.14 d后再行磁共振成像、CT检查,计算肿瘤体积.处死实验兔,免疫组织化学方法检测肿瘤细胞凋亡、血管内皮生长因子及Ⅷ因子表达情况,对Ⅷ因子阳性血管内皮细胞进行微血管密度计数,并结合其病理学特征进行分析.多组间的比较用方差分析,多组数据两两比较用Student-Newwman-Keuls法检验,两组间比较用成组t检验,对方差不齐者用秩和检验.结果 分组处理前,A、B、C、D、E组肿瘤体积分别为(79.4±8.2)、(75.3±7.8)、(74.6±6.6)、(78.7±9.1)、(75.8±8.4)mm3,各组比较,F=12.248,P=0.0636,差异无统计学意义.处理后5组肿瘤体积分别为(564.7±96.7)、(176.5±83.2)、(239.6±42.8),(159.8±58.6)、(334.7±32.6)mm3,各组比较,F=24.537,P=0.0218,差异有统计学意义.处理前后肿瘤增长倍数分别为6.9、2.6、3.1、1.6倍和4.1倍.5组凋亡率分别为12.0%±1.1%、14.5%±2.1%、17.6%±2.3%、18.6%±2.3%和19.6%±2.5%,各组比较,F=6.23,P=0.024,差异有统计学意义.血管内皮生长因子表达阳性率分别为50.0%、83.3%、83.3%、50.0%和50.0%,各组比较,F=7.84,P=0.019,差异有统计学意义.5组吸光度值分别为81.6±16.1、85.2±24.3、75.2±23.9、71.1±21.3和72.3±25.2,各组比较,F=0.854,P=0.018,差异有统计学意义.结论 rAd-p53对兔VX2肝癌有较好的治疗效果,以与超液态碘油经肝动脉联合灌注治疗最为有效.     

Overexpression of p53 and neoplastic cell proliferation in undifferentiated nasopharyngeal carcinoma

Abstract. The author investigated the p53 status in correlation with cellular proliferation in the undifferentiated subgroup, which is infrequently found in caucasians. The author evaluated formalin-fixed, paraffin embedded tissue blocks from sixty cases with undifferentiated carcinoma of the nasopharynx by p53 and Ki67 immunostaining. All samples were retrieved from the surgical pathology file at King Chulalongkorn Memorial Hospital from 2001-2005. The patients had a mean age of 47 years. Stage IV was the most common stage, found in 21 cases (35%). Forty-four tumors (73%) overexpressed p53 protein, which was significantly associated with high rate of tumor cell proliferation (r = 0.477, p < 0.001). The higher the amount of p53 stained, the higher the rate of tumor cell proliferation. However, there was no statistically significant association between p53 protein overexpression and clinical status, including tumor volume, nodal status, and metastatic condition. This observation may explain why some tumors are resistant to radiation and are poorly controlled when they recur in distant organs.     

rAd-p53 enhances the sensitivity of human gastric cancer cells to chemotherapy

AIM:To investigate potential antitumor effects of rAd-p53 by determining if it enhanced sensitivity of gastric cancer cells to chemotherapy.METHODS:Three gastric cancer cell lines with distinct levels of differentiation were treated with various doses of rAd-p53 alone,oxaliplatin(OXA) alone,or a combination of both.Cell growth was assessed with an 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay and the expression levels of p53,Bax and Bcl-2 were determined by immunohistochemistry.The pre...     

Experimental study on ultrasound-guided intratumoral injection of“Star-99”in treatment of hepatocellular carcinoma of nude mice

AIM: To investigate the anti-cancer effect and the immunological mechanism of ultrasound-guided intratumoral injection of Chinese medicine "Star-99" in hepatocellular carcinoma (HCC) of nude mice.METHODS: Twenty-eight human hepatocellular carcinoma SMMC-7721 transplanted nude mice, 14 of hypodermically implanted and 14 of orthotopic liver transplanted, were randomly divided into three groups of which 14 mice with Star-99, and 7 with ethanol and saline respectively. Ten days after the transplantation the medicines were injected into the tumors of all the nude mice once every 5 days.After 4 injections the nude mice were killed. The diameters of three dimension of the tumors were measured by high frequency ultrasound before and after the treatment and the tumor growth indexes* (TGI) were calculated.Radioimmunoassay was used to detect the serum levels of interleukin-2 (IL-2) and tumor necrosis factor (TNF)-alpha.The tumor tissues were sent for flow cytometry (FCM) DNA analysis. Apoptotic cells were visualized by TUNEL assay.All the experiments were carried out by double blind method. zRESULTS: The TGI of Star-99 group (0.076±0.024) was markedly lower than that of the saline group (4.654±1.283)(P＜0.01). It also seemed to be lower than that of the ethanol group (0.082±0.028), but not significantly different (P＞0.05).Serum levels of IL-2 and TNF-α were markedly higher than those of ethanol group and saline groups (P＜0.05). The mean apoptotic index (AI: percentage of TUNEL signal positive cells)in Star-99 group (48.98±5.09 %) was significantly higher than that of the ethanol group (11.95±2.24 %) and the saline group (10.48±3.85 %) (P＜0.01). FCM DNA analysis showed that the appearance rate of the apoptosis peak in Srar-99group was 92.9 %, markedly higher than that of the ethanol group (14.3 %) and the saline group (0.0 %) (P＜0.01).Correlation (r=0.499, P＜0.05) was found between AI and serum level of TNF-α.CONCLUSION: Star-99 has an effect on the elevation of the serum levels of IL-2 and TNF-α. ft indicates that Star99 has the function of enhancing the cellular immunity and inducing cancer cell apoptosis. The correlation between AI and serum level of TNF-α indicates that the elevation of the serum of TNF-α induced by Star-99 may be an important factor in the promotion of the hepatic cancer cell apoptosis.Star-99 has strong effects on the inhibition and destruction of cancer cells. Its curative effect is as good as ethanol. Its major mechanisms can be as follows: (1) it increases the serum levels of IL-2 and TNF-α and triggers cellular immunity. (2) It can induce cancer cells apoptosis, the effective mechanism of the Star-99 is different from that of the ethanol. The mechanisms of triggering the immunologic function of the organism and inducing cell apoptosis are, of particular significance. This study will provide a new pathway of drug administration and an experimental basis for the treatment of HCC with Chinese herbal, and the study of Star-99 in the treatment of tumor is of profound significance with good prospects.     

Gene transfer of somatostatin receptor type 2 by intratumoral injection inhibits established pancreatic carcinoma xenografts

AIM: To investigate the therapeutic effect of somatostatin receptor type 2 (SSTR2) gene transfection on pancreatic carcinoma xenografts in vivo in experimental cancers. METHODS: Human pancreatic cancer cell line Panc-1 was inoculated subcutaneously into the back of nude mice. When tumor nodules were grown as large as about 5 mmx5 mm days after inoculation, the mice were randomly divided into 3 groups (6 mice in each group). Group Ⅰ served as untreated control group. Group Ⅱ received an intratumoral injection of a combination of human cytomegalovirus promoter-6C (pCMV-6C) and lipofectamine 2000. Group Ⅲ received an intratumoral injection of a combination of pCMV-6C-SSTR2 and lipofectamine 2000. The rate of tumor growth was compared among these three groups. The expression of SSTR2 in these tumors was detected by immunohistochemistry and Western-blot. Apoptosis index (AI) in these tumors was examined by using TUNEL in situ. RESULTS: Intratumoral injection of a combination of pCMV-6C-SSTR2 and lipofectamine 2000 resulted in the expression of SSTR2 protein. The tumor size and weight in group Ⅲ (0.318±0.098 cm3, and 0.523±0.090 g, respectively) were significantly lower than those in group I (2.058±0.176 cms, and 1.412±0.146 g, respectively) and group Ⅱ (2.025±0.163 cm3, and 1.365±0.116 g, respectively) (P<0.05) The AI in group Ⅲ (1.47±0.13%) was significantly higher than that in groupⅠ(0.56±0.09%) and group Ⅱ (0.57±0.11%) (P<0.05). But there were no significant differences between groups Ⅰ and Ⅱ. CONCLUSION: Our data demonstrate that re-expression of SSTR2 gene has antitumor effects on experimental pancreatic cancer. Restoration of SSTR2 gene expression through gene transfer in vivo might be a potential gene therapy strategy for human pancreatic cancer.     

An infrequent point mutation of the p53 gene in human nasopharyngeal carcinoma.

Point mutations in the p53 gene have been detected in a variety of human cancers; the mutations are clustered in four "hot-spots" located in the coding region of exons 5, 7, and 8, which coincide with the four most highly conserved regions of the gene. We report the finding of a heterozygous G----C mutation at codon 280 (exon 8), position 2, of the p53 gene in a nasopharyngeal carcinoma (NPC) cell line, originating from Guangdong, a province in the People's Republic of China that leads the world in NPC incidence. A survey of nasopharyngeal tissues and NPC biopsies revealed that 1 out of 12 NPC samples from Hunan, another province in the People's Republic of China with high NPC incidence, had the same heterozygous mutation at codon 280 of p53, and none of 10 biopsies from Taiwan showed a mutation within exons 5-8 of the p53 gene. No other alteration of gene structure, including gross rearrangement or loss of heterozygosity or abnormality of gene expression was detected in NPC cell lines or NPC biopsies. We conclude from this study that mutational or other alterations of the p53 gene are not common in nasopharyngeal carcinogenesis and that a codon-280 mutation of p53 may be involved in less than 10% of NPC cases. This result contrasts with the relatively high frequency of p53 mutations associated with several other human carcinomas and suggests the importance of other genes in NPC genesis.