Effect of polypeptide CH50 on macrophage activationin vivo and antitumor function

Surgery,chemotherapy and radiotherapy aremajor methods used in tumor therapy.After the re-moval of large tumor tissue by surgery,the mainproblem to be faced with is the inhibition and elimi-nation of the residual tumor cells in vivo. Ifmacrophages can be effectively activated,the residualtumor cells could be inhibited and eliminated as earlyas possible,the conditions favorable to the induce-mentof specificimmune response againsttumorcouldalso be created. Recombinant polypeptide CH50 wasprepar…     

Local injection of liposomal adriamycin to inhibit metastatic cell proliferation in axillary nodes in rabbits bearing breast tumors

Objective： To assess the inhibitory effects of local injection of liposomal adriamycin （LADR） on the proliferation of lymph node metastases in rabbits bearing VX2 carcinoma in the mammary gland. Methods：Thirty female New Zealand white rabbits were divided into 3 groups, with 10 in each. VX2 tumor mass suspensions were injected into the breast tissues of rabbits. Treatment initiated once the axillary lymph node reached 5 mm in the maximum diameter. Group 1 received a sham treatment. Group 2 received a subcutaneous injection of LADR adjacent to tumor. Group 3 received an intravenous injection of free ADR （FADR） at the same dose and concentration to group 2. The breast tumors and axillary lymph nodes were resected after the treatment was repeated 3 times. The tumor and node sizes before and after treatment were measured. PCNA mRNA expressions in breast tumors and axillary nodes were determined using RT-PCR. Results： The mean growth ratios of lymph nodes after treatment were 3. 70, 1.55, and 2.89, respectively, in groups 1, 2, and 3. The slowest node growth was observed in animals of group 2, with significant differences from group 1 （P〈0. 001） and group 3 （P= 0. 002）. The relative values of PCNA mRNA expression in lymph nodes were 0. 541, 0. 329, and 0. 450, respectively, in groups 1, 2, and 3. Group 2 exhibited a significantly reduced PCNA mRNA expression in metastatic lymph node, as compared to group 1 （P〈0. 001） and group 3 （P=0. 004）. Intravenous FADR injection effectively lowered the mRNA expressions of PCNA in breast tumors, which were not apparently altered after local LADR injection. Conclusion： Local injection of LADR holds a strong inhibitory effect on the proliferation of metastatic tumor cells in lymph nodes and appears to be an effective method for the treatment of lymphatic metastases of breast cancer.     

银杏叶提取物联合前列腺素E1治疗糖尿病下肢血管病变的研究

目的 探讨银杏叶提取物（EGB）注射液联合前列腺素E1（PGE1）注射液治疗糖尿病下肢血管病变（PAD）的有效性和安全性.方法 将100例糖尿病PAD患者随机分为两组,对照组50例给予PGE1注射液静脉注射治疗;治疗组在此治疗基础上给予EGB注射液静脉注射治疗.2周后比较两组治疗前后临床症状体征、血液流变学相关指标和踝肱指数（ABI）指标的变化.结果 治疗组总有效率为94.0％,对照组总有效率为80.0％,两组治疗结果比较有统计学意义（P＜0.05）;2组血液流变学明显改善,ABI水平显著提高,治疗前后指标比较P＜0.05,治疗组指标改善明显优于对照组,差异均有统计学意义（P＜0.05）.结论 银杏叶提取物注射液联合前列腺素E1注射液能缓解糖尿病下肢血管病变患者临床症状,改善血液流变学和踝肱指数,有显著的临床疗效.     

重组腺相关病毒介导TRAIL对卵巢癌裸鼠肝转移的抑制作用的实验研究

目的探讨重组腺相关病毒介导TRAIL对卵巢癌裸鼠肝转移的抑制作用。方法构建卵巢癌裸鼠肝转移模型。选择重组腺相关病毒rAAV—PEG—sTRAIL治疗，将裸鼠分为TRAIL治疗组和对照组，治疗6w后，观察肝转移率和肝转移结节数；Tunnel法分析TRAIL对肿瘤细胞的凋亡诱导情况。结果全身系统性给予重组腺相关病毒介导TRAIL后，治疗组和对照组相比转移率低，肝转移结节数目少，差异有统计学意义（p〈0．05）；Tunnel法分析TRAIL可诱导肿瘤细胞凋亡。结论重组腺相关病毒介导TRAIL可抑制卵巢癌裸鼠肝的转移生长，TRAIL用于卵巢癌肝转移的基因治疗中具有较好的应用前景。     

肝动脉栓塞化疗联合静脉化疗在胃癌肝转移中的疗效分析

目的探讨肝动脉栓塞化疗联合静脉化疗在胃癌肝转移中的疗效。方法对145例胃癌肝转移瘤行肝动脉造影,观察肝转移瘤血供情况,进行肝动脉栓塞化疗,并联合静脉化疗,评价其疗效。结果胃癌肝转移瘤血供丰富86例,有效率88.37%,中位生存期16.5个月;胃癌肝转移瘤少血供59例,有效率49.42%,中位生存期9.2个月。两组间有明显差异（P〈0.05）。结论肝动脉栓塞化疗联合静脉化疗是胃癌肝转移的有效治疗方法,富血供的胃癌肝转移瘤较少血供的胃癌肝转移瘤疗效好。     

The Expression and Activity of MMPs Are Increased in Residual Tumor Tissues after the Termination of Immunotherapy

To investigate the invasive ability of the residual tumor cells after immunotherapy and explore the feasible approach suppressing the invasion, mice were inoculated with B16 cells, and then treated by gene therapy with p4-1BBL/psPD-1 or IFN-γ. The production and activities of MMP-9 and MMP-2 in residual tumor tissues were analyzed with gelatin zymography 1 day and 7 days after the termination of the immunotherapy. The production of MMP-9 and MMP-2 by B16 cells treated with IFN-γ was also analyzed. IFN-γ-treated B16 cells were inoculated to mice via subcutaneous injection. The invasion of tumor to muscular tissue was analyzed. Gene therapy with CH50 was used to suppress the invasive growth of tumor. The results showed that the expression and the activities of MMP-9 and MMP-2 were significantly increased 7 days after the end of immunotherapy. The re- sponse of tumor cells to ECM molecules was intensified after the removal of IFN-γ, resulting in significant increase of both the production and activities of MMP-9 and MMP-2, and the increased invasion of tumor. Gene therapy with CH50 effectively suppressed the invasive growth of tumor. It is concluded that the termination of immunotherapy may result in a higher metastatic potential of residual tumor cells. Suppressing tumor invasion by suitable treatment will improve the efficacy of immunotherapy..     

重组FN多肽CH50及IFN-γ对化疗小鼠免疫功能影响的比较

腹腔连续注射ＣＨ５０或腹腔内转染ＩＦＮ－γ基因能减弱或防止化疗药物对免疫功能的抑制作用,减轻或防止化疗药物使外周血单核细胞数量和腹腔细胞的数量降低（Ｐ＜０．０５）,促进巨噬细胞代谢活性、杀瘤活性以及分泌细胞因子的能力（Ｐ＜０．０５,＜０．０１）;促进脾淋巴细胞的增殖反应能力（Ｐ＜０．０５,＜０．０１）。注射ＣＨ５０的作用早于ＩＦＮ－γ基因转染的作用。结果提示ＣＨ５０可改善肿瘤化疗药物疗效。     

人胃癌裸鼠原位移植和转移模型的建立及三种方法比较

目的：建立胃癌裸鼠原位移植和转移模型，并比较三种不同方法的结果。方法：将胃癌肿瘤细胞悬液、胃癌裸鼠皮下移植瘤块和胃癌裸鼠原位移植瘤块种植于裸鼠胃壁，形成原位移植瘤，观察和比较三种方法所建立的模型肿瘤生长状况、移植成功率和自发转移的发生率。常规HE染色，观察胃癌原位移植瘤、胃癌淋巴结转移和肝转移的病理切片。结果：细胞悬液种植的原位胃癌成瘤率为50％，淋巴结广泛转移率为37．5％，肝转移发生率为25％；皮下瘤块的原位成瘤率为100％，淋巴结广泛转移率为62．5％，肝转移发生率为50％，腹水形成率为6．25％；而胃癌裸鼠原位移植块再次原位移植的原位成瘤率达100％，淋巴结广泛转移率为87．5％，肝转移发生率为75％，腹水形成率为12．5％。细胞悬液成瘤时间较皮下肿瘤组织块和胃癌裸鼠原位移植块再次移植延迟2—3周，而后两者无差异。结论：三种原位移植和转移模型均具有人胃癌自然生长过程的特点，以胃癌裸鼠原位移植块再次移植为最优，且更具有人胃癌的生物学活性。     

Construction of eukaryotic expressing vector pCH503 of CH50 and its chemotaxis and anti-tumor function by expressionin vivo in mice

Summary An eukaryotic expressing vector that expresses CH50, a recombinant polypeptide of human fibronectin, in mice was constructed, and its chemotactic and anti-tumor function byin vivo gene transfection was investigated. The plasmid was constructed by recombination techniques. The cDNA fragment coding CH50 polypeptide from a prokaryotic expressing vector of CH50 was ligated with 5′-terminal noncoding region and coding region of signal peptide of mouse IFN-7 cDNA at 5′ side and 3′-terminal noncoden region of human FN cDNA at 3′ side. The recombinant cDNA was inserted into plasmid pREP8. The resulted expressing plasmid was designated as pCH503. The macrophages transfected with pCH503in vivo and culturedin vitro could produce CH50. The expressed product was identified by heparin-affinity chromatography and SDS-PAGE. By counting and Giemsa-staining of coeliac cells and histotomy and staining of muscle tissue, the chemotaxis on immune cells was observed after transfection of pCH503 either in peritoneal cavity or in muscle. The inhibition of gene transfection of pCH503 on melanoma was observed in mice. The number of melanoma nodes in mice was reduced by 50%–60% after coeliac transfection with pCH503. The pCH503, an eukaryotic expressing vector of CH50, can expressin vivo in mice. The transfection of pCH503in vivo has the chemotaxis on immune cells and can inhibit the formation of tumor nodes, suggesting that plasmid pCH503 is potentially useful in combined treatment of tumor. This project was supported by a grant from the National Natural Science Foundation of China (No. 39870763) and Trans-Century Training Program Foundation for Talents under the Supervision of Ministry of Education of China.     

人胃癌裸鼠原位移植和转移模型的建立及两种方法比较

目的：建立人胃癌裸小鼠原位移植和转移模型，并比较两种不同方法的结果。方法：将肿瘤组织块和肿瘤细胞悬液种植于裸小鼠胃壁形成原位移植瘤，观察和比较了两种方法所建立模型肿瘤生长状况、移植成功率和自发转移的发生率。结果：胃癌组织块种植的原位成瘤率达100％，淋巴结广泛转移率90％，肝转移发生率75％。细胞悬液种植的原位成瘤时间较组织块法延迟2周，成瘤率为67％，淋巴结广泛转移率为50％，肝转移发生率仅为30％。结论：两种原位移植和转移模型均具有人胃癌自然生长过程的特点，以肿瘤组织块原位模型为优，该模型的建立可为人胃癌转移机制和抗转移实验治疗的研究提供一种有价值的工具。     

Construction and Expression of Eukaryotic Expressing Vector pCH510 of Polypeptide CH50 and Its Chemotaxis and Antitumor Function by in vivo Transfection

Fibronectin ( FN) is a high- molecular glycopro-tein with multiple- domain.FN fragments containingCell domain are chemotactic to macrophages[1] .CH5 0 polypeptide,a recombinant polypeptide weproduced by the recombination of Cell and Hep domains of human fibronectin and expression in E.coli,can activate macrophages and inhibitthe metas-tasis of tumors[2 -4 ] .To investigate the function ofCH5 0 expressed in vivo by gene transfection,explorethe feasibility of applying the polypeptide to gene…     

健脾补肾泄浊法治疗肾性贫血30例临床观察

目的:观察健脾补肾泄浊法配合促红细胞生成素(EPO)治疗肾性贫血的临床疗效。方法:治疗组给予EPO(宁红欣)3000u皮下注射,并口服健脾补肾泄浊中药;对照组只给予EPO3000u皮下注射。结果:治疗组总有效率为86.2%,对照组总有效率为70.0%。治疗组疗效优于对照组,差异有显著性(P<0.05)。两组治疗后血红蛋白、红细胞比容均较治疗前明显上升(P<0.01),但治疗组优于对照组(P<0.05);治疗组血清肌酐、尿素氮、血清总蛋白治疗后与治疗前相比,差异有显著性(P<0.05),两组间比较,差异亦有显著性(P<0.05)。结论:健脾补肾泄浊法配合EPO治疗肾性贫血,能明显改善患者贫血状况,对肾功能有较好的保护作用,提高患者生活质量。     

来源于HHV8 MIP N端多肽抗乳腺癌作用的实验研究

目的:探讨来源于HHV8 MIP N端多肽(NT21MP)治疗小鼠乳腺癌的疗效。方法:采用乳腺癌细胞株4T-1构建乳腺癌小鼠模型;实验分为NT21MP5μg/kg、50μg/kg和500μg/kg组,NT21MP与Herceptin联合用约组(NT21MP50μg/kg+Herceptin 36.04μg/kg)、阳性对照组(Herceptin 36.04μg/kg)及生理盐水对照组;观察各组小鼠肿瘤体积大小,并计算抑瘤率;检测肺转移结节。结果:与生理盐水组荷瘤小鼠比较,NT21MP呈剂量依赖性地抑制肿瘤的生长,以联合用药组为明显(P<0.01)。NT21MP不同浓度、联合用药及阳性对照组的抑瘤率分别为10.0%、41.6%、81.0%、58.2%及39.2%;对照组小鼠肺内均见转移性Lewis肺癌瘤灶,肺脏表面可见多个肿瘤转移结节;NT21MP 5μg/kg组小鼠肺内均见转移性Lewis肺癌瘤灶,肺脏表面可见到散在的肿瘤转移结节;NT21MP 50μg/kg组和Herceptin组中,各有5/6动物肺内见转移性Lewis肺癌瘤灶,肺表面可见小的单个肿瘤转移结节;NT21MP 500μg/kg组和联合用药组各有2/6和3/6动物肺内见转移性Lewis肺癌瘤灶,肺表面未见明显的转移结节。结论:NT21MP可抑制乳腺癌的生长和转移,联合应用基因靶向药物Herceptin,可提高对乳腺癌靶向治疗的效果。     

β1整合素反义寡核苷酸联合顺铂治疗人卵巢癌裸鼠移植瘤的研究

目的：探讨以β1整合素反义寡核苷酸（ASODN）治疗人卵巢癌裸鼠皮下移植瘤的可行性.方法：常规体外培养SKOV3细胞,采用皮下注射法建立移植瘤裸鼠动物模型.32只荷瘤鼠随机分为ASODN组（A组）、ASODN联合顺铂（DDP）组（A＋D组）、DDP组和0.9％氯化钠注射液对照组（NS组）,每组8只,分组给药.以脂质体包裹的β1整合素ASODN直接移植瘤内注射,观察肿瘤生长情况,测瘤体积并计算抑瘤率.采用反转录-聚合酶链反应（RT-PCR）和免疫组织化学方法分别检测β1整合素mRNA的表达.结果：A组肿瘤体积和抑瘤率分别为（316.10 ±21.77） mm3和48.15％,与NS组比较抑瘤率较高,肿瘤生长缓慢（P＜0.01）.而A＋D组肿瘤体积和抑瘤率分别为（178.70±40.67） mm3和70.37％,与DDP组、A组及NS组差异均有统计学意义（P ＜0.05 ～P＜0.01）.RT-PCR和免疫组织化学检测,A组和A＋D组肿瘤组织中β1整合素mRNA的表达均明显下调（P＜0.01）.结论：单用β1整合素ASDON或联用DDP均可有效抑制人卵巢癌裸鼠皮下移植瘤组织的生长,可能与其特异性下调β1整合素基因表达有关.特异性靶向β1整合素ASODN可用于卵巢癌的辅助治疗.     

Augmentation of Recombinant Fibronectin Polypeptide CH50 on the Antitumor Function of Macrophages

Fibronectin(FN)isamulti-domainglycoproteinwithcomplicatedregulatoryfunctionsllj.TheapplicationofFNtothetherapyofdiseaseshasbeentriedsince1983,buthampered'becauseofthecomplexityofitsfunctionslll.Inrecentyears,theresearchonFNwasconcentratedonthefunctionsofitsdomainfragments.IthasbeenprovedthattheFNfragmentcontainingCellIdomaincouldenhancethefunctionofmonocyte--macrophage['--'J,suggestingthatdomainfragmentofFNcouldbeveryusefulinthetherapyofdiseases.WehaverecombinedcDNAfragmentsofCellIdoma…     

Construction and expression of eukaryotic expressing vector pCH510 of polypeptide CH50 and its chemotaxis and antitumor function byin vivo transfection

Summary  To construct an eukaryotic expressing vector that expresses CH50, a recombinant Cell I-Hep I bifunctional-domain polypeptide of human fibronectin, and to investigate the chemotaxis to immune cells and the inhibitory effect on the growth of tumor by the expression of the plasmidin vivo, the plasmid was constructed by DNA recombination. Gene transfection was performedin vitro andin vivo. The expressed product was identified by Western blot. The chemotaxis after gene transfectionin vivo was observed by histotomy and staining of muscle tissues. The inhibition of gene transfection on solid tumor was observed in mice. The results showed that plasmid pCH510 was constructed by the recombination of the 5′-terminal noncoding region and signal peptide coding region of human fibronectin cDNA and cDNA fragment coding CH50 polypeptide with a 3′-terminal noncoding region of human FN cDNA, and the insertion of the recombinated fragment into plasmid pcDNA3. 1. After transfection with plasmid pCH510, NIH3T3 cells could produce CH50 polypeptide. The transfection of plasmid pCH510 by the injection in muscle of mouse could produce the effects of chemotaxis on immune cells and the inhibition on the growth of solid tumor. It is concluded that plasmid pCH510 can express in cells andin vivo in mouse. The expression of the plasmidin vivo has a chemotactic effect on immune cells and can inhibit the growth of solid tumor. This project was supported by a grant from the National Natural Science Foundation of China (No. 39870763) and a Funding Program for New-Century Talent of the Ministry of Education of China.     

5-氟脲嘧啶治疗前列腺癌的实验研究

目的 :研究 5-氟脲嘧啶不同给药方式的药物疗效 ,探讨提高其疗效的可行方案。方法 :以前列腺癌肿瘤细胞系 PC- 3M为研究对象 ,观察 5-氟脲嘧啶对 PC- 3M体外细胞系及裸鼠体内肿瘤生长的抑制作用及其毒副作用。结果 :5-氟脲嘧啶对体外肿瘤生长的抑制作用起效浓度1 μg/ml;小剂量 5-氟脲嘧啶 (2 0 mg/kg)肿瘤内注射给药对裸鼠体内移植瘤的抑瘤率为 1 0 .5% ,体重较对照组无减轻 ;大剂量 5-氟脲嘧啶 (50 mg/kg)肿瘤内注射抑瘤率为 71 % ,体重一过性减轻 ;大剂量 5-氟脲嘧啶 (50 mg/kg)全身用药 ,实验动物体重迅速减轻 ,一周内全部死亡。结论 :高浓度 5-氟脲嘧啶有较好的肿瘤抑制作用 ,肿瘤内注射给药可保证局部 5-氟脲嘧啶的高浓度而全身毒副作用较低 ,对于局部复发的前列腺癌有一定的治疗作用 ,并可作为初治病人内分泌治疗的辅助手段     

唑来膦酸治疗骨转移肿瘤疼痛的临床观察

目的评价注射用唑来膦酸治疗骨转移肿瘤疼痛的临床疗效和安全性。方法开放性临床随机选择骨转移肿瘤患者26例,男性17例,女性9例。静脉注射唑来膦酸4 mg,每月一次,1个月为1个周期,2个周期后评价疗效及安全性。疗效评价指标包括止痛疗效、KPS评分的变化和转移灶治疗情况。结果 26例患者中,疼痛完全缓解10例,部分缓解13例,轻微缓解2例,无缓解1例,有效率为88.5%,治疗前KPS评分中位60分,治疗后KPS评分中位70分,治疗后较治疗前提高10分,生活质量明显提高。不良事件的发生率为7.7%,未出现严重不良事件,不良反应为轻度发热、消化道反应等。结论唑来膦酸对骨转移肿瘤引起的疼痛有良好的止痛效果,对转移灶有较好的治疗作用,并且安全性评估结果也比较好,值得临床推广应用。     

CH50多肽对IFN-γ基因转染癌细胞体内生长及免疫刺激作用的影响

探讨重组FN多肽CH50对IFN-γ基因转染癌细胞体内生长及免疫刺激作用的影响。将小鼠IFN-γ基因转染黑色素瘤B16/Fl细胞,测定其表达产物与CH50协同刺激巨噬细胞产生NO的作用、转染细胞接种小鼠并注射CH50时对脾细胞的免疫刺激作用及瘤细胞的体内生长特性。结果表明,转染细胞表达产物可与CH50协同作用刺激巨噬细胞产生NO。转染细胞表达IFN-γ水平较低,仍可在体内形成肿瘤,注射CH50能够抑制其形成肿瘤。基因转染细胞和CH50能够促进脾细胞对肿瘤细胞的杀伤能力。研究表明,CH50不仅可以提高肿瘤疫苗的安全性,也可以提高肿瘤疫苗刺激机体免疫系统的作用。并提示,表达CH50和IFN-γ双因子的肿瘤疫苗可为提高肿瘤疫苗治疗肿瘤的效果开辟新的途径。