H9N2禽流感病毒血凝素和神经氨酸酶DNA疫苗对小鼠的免疫保护研究

目的 检测H9N2禽流感病毒血凝素(hemagglutinin,HA)和神经氨酸酶(neuraminidase,NA)DNA疫苗保护小鼠抵抗致死量同源病毒感染的能力.方法 通过小鼠肺对肺传代,建立禽流感病毒A/chicken/Jiangsu/07/2002(H9N2)小鼠适应株.同时,构建病毒HA和NA DNA疫苗,以不同剂量电击法免疫小鼠1或2次,在初次免疫后4周或加强免疫后1周用致死量(40 LD50)鼠适应型病毒攻击小鼠.通过测定小鼠血清抗体滴度、小鼠存活率和肺部病毒滴度来评价疫占的效果.结果 HA或NADNA 10μg免疫1次或3μg免疫2次均可保护小鼠抵抗致死昔H9N2病毒的感染.结论 低剂量HA或NA DNA可为抗H9N2禽流感病毒感染提供有效的免疫保护.     

禽流感H5N1灭活疫苗与不同纳米化佐剂联合免疫研究

目的 观察禽流感H5N1灭活疫苗加不同佐剂以及纳米化佐剂免疫小鼠后产生的免疫应答的差异,同时观察免疫对异亚型病毒攻击后的保护情况.   方法 用H5N1灭活疫苗分别联合佐剂氢氧化铝、纳米化氢氧化铝、MF59和纳米化MF59通过腹腔注射方式免疫雌性BALB/c小鼠,同时分别以H5N1灭活疫苗免疫小鼠以及PBS腹腔注射处理小鼠作对照.采用ELISA方法分别对各组小鼠免疫后血清特异性IgG及其亚类IgG1、IgG2a水平进行检测,以PR8病毒鼻腔攻击后观察小鼠体重变化情况和生存率.采用t检验作组间比较.    结果 与PBS处理组相比,无论以何种方式免疫H5N1疫苗,免疫后特异性IgG及其亚类水平均明显升高(t=7.4004,P<0.01),以联合MF59后诱导的特异性抗体水平最高.其中疫苗单独免疫组和联合M59免疫组IgG2a水平升高明显,联合纳米化MF59免疫小鼠后IgG2a水平有所下降,IgG1水平有所升高;联合铝佐剂组以IgG1升高为主.攻毒后各组小鼠体重均出现下降,但疫苗单独免疫小鼠以及疫苗与佐剂联合免疫小鼠于攻毒后期体重恢复接近正常或者正常.PBS处理组小鼠攻毒后全部死亡,佐剂联合免疫组小鼠存活率100%,而疫苗单独免疫小鼠存活率为70%.   结论 H5N1疫苗无论是单独免疫或是联合不同佐剂免疫均可诱导较高水平的特异性抗体产生,有非常好的免疫原性.H5N1灭活疫苗诱导的抗体亚类以IgG2a为主,MF59以及纳米化MF59也以诱导IgG2a亚类为主,但纳米化MF59可诱导更均衡的免疫应答.联合铝佐剂或纳米化铝佐剂免疫小鼠后均可诱导以IgG1亚类为主的抗体应答.联合两种佐剂均可对小鼠产生很好的异亚型保护.     

新型冠状病毒受体结合域-Fc融合蛋白表达及小鼠免疫原性

目的  表达制备新型冠状病毒刺突蛋白的受体结合域（receptor binding domain,RBD）-Fc融合蛋白,并评价其在小鼠模型中的免疫原性。方法  将新型冠状病毒RBD与小鼠免疫球蛋白Fc段融合基因在中国仓鼠卵巢细胞中融合表达并制备融合蛋白。将不同剂量的RBD-Fc融合蛋白分别单独或辅以氢氧化铝佐剂免疫小鼠,通过ELISA、假病毒中和试验、酶联免疫斑点试验检测诱导产生的体液和细胞免疫应答。结果  10 μg RBD-Fc融合蛋白辅以氢氧化铝佐剂能有效诱导小鼠产生良好的RBD特异性IgG抗体应答,该抗体可阻断病毒RBD与细胞表面病毒受体的结合,进一步研究表明该重组蛋白还诱导产生了针对原始毒株、Beta变异株和Delta变异株假病毒的中和抗体,中和抗体滴度分别为1 566、336和270。结论  制备的RBD-Fc融合蛋白具有较好的免疫原性,可为开发COVID-19重组蛋白疫苗提供参考。     

对禽流感H5N1灭活疫苗不同免疫方式诱导的免疫应答及异亚型保护研究

目的 观察禽流感H5N1灭活疫苗以不同方式免疫后诱导的免疫应答及抗异亚型病毒攻击的保护作用.方法 将H5N1灭活疫苗分别通过腹腔和滴鼻方式免疫BALB/c小鼠,同时以PBS作为对照;免疫后分别以PR8和H9N2病毒攻击,观察小鼠的体重变化和生存情况.采用ELISA对各组小鼠攻毒后不同时间的血清IgG及其亚类水平进行动态检测;流式细胞仪检测脾淋巴细胞亚群情况.采用t检验对各组数据进行比较.结果 PR8和H9N2病毒攻击后,各组小鼠体重均下降,但疫苗组小鼠于后期体重恢复正常,存活率分别为100%和70%-80%,而PBS组小鼠则全部死亡.无论以何种方式免疫,疫苗组的特异性IgG及其亚类水平均明显升高,其中以IgG2a水平升高更为明显.攻毒后疫苗组小鼠脾CD4+与CD8+T淋巴细胞比值出现下降(t=6.8017,P<0.01);滴鼻免疫组与腹腔免疫组相比降低更明显(t=3.9701,P<0.05).结论 H5N1疫苗免疫原性良好,可诱导较高水平的特异性抗体产生,诱导的抗体亚类以IgG2a为主.两种免疫方式均可对小鼠提供很好的异亚型保护,而滴鼻免疫能够诱导更强的CD8+T细胞应答.

Abstract：

Objective To observe immune responses and heterosubtypic protection elicited by an inactivated influenza H5N1 vaccine with different immunization routes in mice. Methods BALB/c mice were intraperitoneally injected or intranasally immunized with the inactivated H5N1 vaccine, the mice administered with PBS were used as control. Weight loss and survival in mice were observed after PR8 or a H9N2 virus challenge. Serum specific IgG antibody and its subclasses were detected by ELISA kits. Ratios of CD4+ /CD8+ lymphocytes in spleens of mice were assayed. The t-test was used in the comparison of different groups.Results After challenge, weight loss was found in all groups, but the weight of mice in vaccination groups returned to normal later. The mice in PBS groups all died. The vaccinated mice were completely protected against PR8 and partly protected against H9N2 virus. The level of IgG antibody increased significantly after vaccination, and the increase magnitude of IgG2a was higher than that of IgG1. The ratios of CD4+ /CD8+ lymphocytes in spleens of vaccinated mice decreased after challenge (t=6. 8017,P<0. 01) , especially in the intranasal group (t = 3. 9701, P < 0. 05 ). Conclusions Both intraperitoneal injection and intranasal immunization can induce a high level of specific IgG, especially the level of IgG2a, and provide protection against heterosubtypic viruses. Intranasal immunization seems to induce a higher level of CD8+ T cell response.     

鹿茸多肽对阿霉素诱导H9c2细胞损伤保护作用的研究

目的：探讨鹿茸多肽对阿霉素诱导的H9c2细胞损伤的保护作用与其作用机制。方法：培养H9c2细胞，采用CCK-8方法考察ADR和PAP对H9c2细胞存活率的影响，选择最优给药时间与浓度，分成4组，空白对照组（BCG组）、阿霉素诱导组（ADR组）、鹿茸多肽组（PAP组）、阿霉素与鹿茸多肽联合诱导组（ADR+PAP组），ELISA检测H9c2上清液中cTnT和cTnI的含量，免疫荧光法检测Caspase9的表达水平，Western blot法检测各组细胞相关蛋白TGF-β₁、SMAD7、PKC表达。结果：与BCG组比较，ADR组cTnT和cTnI含量显著升高（P<0.05），Caspase9活性表达降低，TGF-β₁蛋白表达升高（P<0.01），SMAD7、PKC蛋白表达降低（P<0.01），PAP组无显著性差异（P>0.05），与ADR组比较，ADR+PAP组cTnT和cTnI含量减少（P<0.05），Caspase9活性表达升高，TGF-β₁蛋白表达降低（P<0.05），SMAD7、PKC蛋白表达升高（P<0.01，P<0.05）。结论：鹿茸多肽可以对阿霉素诱导的H9c2细胞损伤有保护作用，其作用机制可能与调控TGF-β₁、SMAD7、PKC蛋白表达量有关。     

表达甲型H1N1流感病毒血凝素抗原的DNA疫苗构建及其免疫原性评价

目的 构建表达甲型H1N1流感病毒血凝素(hemagglutinin,HA)抗原的DNA疫苗,并在小鼠中测试其免疫原性.方法 运用人密码子优化技术合成甲型H1N1流感病毒HA序列,并转移至DNA疫苗载体,用Western blotting检测其表达效率.采用单纯随机抽样方法把BALB/c小鼠分成DNA疫苗组和对照组.用...     

对禽流感H5N1灭活疫苗与不同免疫方式诱导的免疫应答及异亚型保护研究

目的  观察禽流感H5N1灭活疫苗以不同方式免疫后诱导的免疫应答及抗异亚型病毒攻击的保护作用.  方法 将H5N1灭活疫苗分别通过腹腔和滴鼻方式免疫BALB/c小鼠,同时以PBS作为对照;免疫后分别以PR8和H9N2病毒攻击,观察小鼠的体重变化和生存情况.采用ELISA对各组小鼠攻毒后不同时间的血清IgG及其亚类水平进行动态检测;流式细胞仪检测脾淋巴细胞亚群情况.采用t检验对各组数据进行比较.   结果 PR8和H9N2病毒攻击后,各组小鼠体重均下降,但疫苗组小鼠于后期体重恢复正常,存活率分别为100%和70%-80%,而PBS组小鼠则全部死亡.无论以何种方式免疫,疫苗组的特异性IgG及其亚类水平均明显升高,其中以IgG2a水平升高更为明显.攻毒后疫苗组小鼠脾CD4+与CD8+T淋巴细胞比值出现下降(t=6.8017,P<0.01);滴鼻免疫组与腹腔免疫组相比降低更明显(t=3.9701,P<0.05).    结论   H5N1疫苗免疫原性良好,可诱导较高水平的特异性抗体产生,诱导的抗体亚类以IgG2a为主.两种免疫方式均可对小鼠提供很好的异亚型保护,而滴鼻免疫能够诱导更强的CD8+T细胞应答.     

牛至油增强小鼠免疫功能的实验

目的：研究牛至油对小鼠免疫功能的影响。方法：采用ConA诱导的小鼠脾淋巴细胞转化实验（MTT法）、NK细胞活性测定、小鼠腹腔巨噬细胞吞噬鸡红细胞实验等方法考察牛至油对小鼠免疫功能的作用。结果：牛至油在促进NK细胞活性和单核-巨噬细胞方面的具有明显的作用，但在ConA诱导的小鼠脾淋巴细胞转化能力试验中并未表现出显著的差异。结论：牛至油确实具有增强机体免疫功能的作用。     

人甲型H7N9禽流感全病毒灭活疫苗和裂解疫苗在小鼠中的免疫原性研究

 目的   比较人甲型H7N9禽流感全病毒灭活疫苗和裂解疫苗在小鼠中的免疫原性,为该疫苗的类型选择提供初步依据。 方法   采用相同血凝素含量（5 μg）的H7N9全病毒灭活和裂解疫苗（含或不含氢氧化铝佐剂共4种类型）,分别对BALB/c小鼠进行1针或2针免疫。免疫后,用血凝抑制（hemagglutination inhibition,HI）试验检测血清抗体滴度,比较不同类型疫苗的免疫效果。 结果   小鼠免疫1针全病毒灭活疫苗后,全部血清阳转,HI抗体几何平均滴度（geometric mean titer,GMT）为149;免疫2针后,抗体GMT为243。小鼠免疫1针裂解疫苗后无抗体阳转;免疫2针后全部抗体阳转,GMT为139。两种疫苗添加铝佐剂后,诱导的HI抗体GMT仅略有增加。 结论   H7N9全病毒灭活疫苗在小鼠中的免疫原性较强。在同样类型和剂量的情况下,裂解疫苗需要免疫两次才能达到与全病毒疫苗相同的效果。铝佐剂对免疫原性提升不明显。     

抗H7N9流感病毒中和抗体快速检测方法的建立及应用

目的：对建立的抗H7N9流感病毒中和抗体快速检测方法进行方法学验证及初步应用。方法：分别采用不同代次细胞对高、中、低不同滴度的阳性血清进行多次平行检测,考察细胞代次对检验结果的影响;采用NIBSC提供的参考品对方法学的特异性进行验证;同时应用抗H7N9的阳性血清检测进一步评估方法的准确性和精密性;采用ELISA-MNT法和血凝抑制（hemagglutination inhibition,HI）试验分别接种H7N9灭活流感疫苗免疫的小鼠血清样本20份,分析两种方法检测结果的相关性。结果：采用ELISA-MNT中和法,使用不同代次MDCK细胞（25、30和35代）检测相同的血清样本的中和抗体滴度结果相同;该方法只对羊抗H7N9的血清具有较高保护力,对其他血清基本没有交叉反应;该方法准确性良好;该方法组间、组内精密性的平均变异系数分别为4%和11%。该方法测定H7N9型流感疫苗免疫后的小鼠血清抗体效价,其结果与HI抗体的相关系数为0.61,表明两种方法的检测结果之间呈良好的正相关性。结论：建立的微量病毒中和法能够满足H7N9流感病毒中和抗体效价检测的要求,可用于H7N9新型大流行流感疫苗的免疫评价。     

人用H5N1禽流感病毒疫苗生产工艺的优化

目的 建立优化的人用H5N1禽流感病毒疫苗生产的工艺。方法 在不同的稀释倍数、收获时间及灭活剂添加量下,通过测量收获液的病毒滴度和血凝效价,来确定病毒的最佳生产条件,并对离心法和凝胶过滤层析法的纯化效果进行对比。结果 103~104半数鸡胚感染量(50% egg infective dose,EID50)病毒接种鸡胚,收获的鸡胚尿囊液的病毒滴度和血凝效价最高,分别为10-8.3EID50和1∶480;在56~72 h血凝效价最高。甲醛浓度1∶10 000灭活144 h为灭活最佳条件。两种纯化方法得到的样品纯度和卵清蛋白的去除率相近,但离心纯化法和凝胶过滤层析纯化法病毒回收率有较大的差异,分别为19%和70%。结论 成功建立了高产毒的鸡胚基质H5N1禽流感病毒培养、灭活及纯化工艺。     

Cationic supported lipid bilayers for antigen presentation

Polystyrene sulfate (PSS) particles (301 nm mean diameter) were covered with single cationic dioctadecyldimethylammonium bromide (DDA) bilayers and used for antigen adsorption and presentation. The antigen was a mixture of purified 18/14 Taenia crassiceps proteins (18/14-Tcra). Firstly, the DDA/PSS assembly was characterized at 1 mM NaCl and 5 × 10⁹ PSS particles/mL over a range of DDA concentrations (0.001–1 mM) by means of dynamic light scattering for particle sizing and zeta-potential analysis. 0.01 mM DDA is enough to produce homodisperse and cationic bilayer-covered particles. Secondly, under these experimental conditions, 18/14-Tcra adsorption isotherms onto biomimetic particles or aluminium hydroxide (Al(OH)₃) yield limiting adsorption of 0.36 and 1.32 mg protein/mg biomimetic particles or Al(OH)₃, respectively. Finally, in mice, superior humoral and cellular immunoresponse from serum IgG and footpad swelling was obtained for antigen/biomimetic particles in comparison to conventional Al(OH)₃. Cationic bilayer-covered particles are a novel, highly organized and, possibly, general immunoadjuvant for antigen presentation and subunit vaccine design.     

Long-lasting protective immunity against H7N9 infection is induced by intramuscular or CpG-adjuvanted intranasal immunization with the split H7N9 vaccine

There is an urgent need for efficient vaccines against the highly pathogenic avian influenza A viral strain H7N9. The duration and intensity of the immune response to H7N9 critically impacts the epidemiology of influenza viral infection at the population level. However, the insufficient immunogenicity of H7N9 raises concerns about vaccine efficacy. In this study, we evaluated the impact of immunization routes and the adjuvant CpG on the immune response to a split H7N9 vaccine in mice. Determination of humoral and cellular responses to the vaccine revealed that after four vaccine doses, high titers of H7N9-specific serum IgG, determined by the influenza hemagglutination inhibition (HI) assay, were induced through the intramuscular (i.m.) route and lasted for at least 40 weeks. CpG-adjuvanted immunization increased the levels of long-lived IFN-γ⁺ T cells and raised the Th1-biased IgG2a/IgG1 response ratio. In addition, aside from mucosal IgA, CpG-adjuvanted intranasal (i.n.) immunization elicited serum IgG and cellular responses of a similar duration and intensity to CpG-adjuvanted i.m. immunization. Mouse challenge assays demonstrated that 24 weeks following i.m. immunization without CpG or CpG-adjuvanted immunization through the i.m. or i.n. routes, both offered a high level of protection against H7N9 infection. These results indicate that efficient long-term protection against H7N9 can be achieved via the optimization of vaccination strategies, such as immunization doses, routes, and adjuvants.     

Effects of isoquinoline derivatives, HA1077 and H-7, on cytosolic Ca level and contraction in vascular smooth muscle

The effects of isoquinoline derivatives, HA1077 (1-[5-isoquinolinesulfonyl]-homopiperazine) and H-7 (1-[5-isoquinoline-sulfonyl]-2-methylpiperazine), on cytosolic Ca²⁺ levels ([Ca²⁺]_i) and muscle tension were examined in vascular smooth muscle of rat aorta. High K⁺ (72.7 mM) and norepinephrine (1 μM) induced a sustained contraction with a sustained increase in [Ca²⁺]_i. HA1077 and H-7 (3–10 μM) inhibited the increse in muscle tension more strongly than the increase in [Ca²⁺]_i. Verapamil (10 μM) completely inhibited the increase in [Ca²⁺]_i and the contraction induced by K⁺ whereas it inhibited the increase in [Ca²⁺]_i more strongly than the contraction due to norepinephrine. The verapamil-insensitive portion of the norepinephrine-induced contraction was inhibited by HA1077 or H-7. In Ca²⁺-free solution, 0.1 μM norepinephrine induced a transient increase in [Ca²⁺]_i and muscle tension. The transient contraction was inhibited by 10 μM HA1077 or 10 μM H-7 without inhibiting the increase in [Ca²⁺]_i. 12-Deoxyphorbol 13-isobutyrate (DPB) (1 μM) caused a sustained contraction, and this contraction was inhibited by HA1077 and H-7 at similar concentrations needed to inhibit the contractions induced by high K⁺ or norepinephrine. In rabbit mesenteric artery permeabilized with Staphylococcus aureus -toxin, 100 μM HA1077 and 100 μM H-7 inhibited the contraction induced by 0.3 μM Ca²⁺. These results suggest that the inhibitory effects of isoquinoline derivatives, HA1077 and H-7, are due to a decrease in [Ca²⁺]_i and in the Ca²⁺ sensitivity of contractile elemenst in vascular smooth muscle.     

高致病性H7N9禽流感病毒研究进展

摘要：H7N9禽流感病毒在中国出现以来共造成5次流行。在第5次流行中出现了高致病性H7N9变异株,该病 毒株的HA链接肽位置发生了基因插入性突变,导致该病毒对家禽毒力的增强。同时在人感染H7N9禽流感病例中 也相继分离到了该病毒。因此,对高致病性H7N9禽流感病毒病原学及流行病学研究对于该疾病的预防和控制具有 重要意义。本文从高致病性H7N9禽流感病毒的变异来源、流行病学特征及防治措施等方面进行综述,为高致病性 H7N9禽流感的有效防治提供科学策略。     

流感疫苗病毒株在MDCK细胞中的培养条件

 目的   研究4价流感疫苗所用病毒株在MDCK细胞中的扩增条件。方法 在6孔细胞培养板中以不同感染复数（multiplicity of infection,MOI）（0.100 0、0.010 0、0.001 0、0.000 1）和对甲苯磺酰-L-苯丙氨酸氯甲基酮（TPCK）-胰蛋白酶（胰酶）浓度（0、2、4、8 μg/ml）进行病毒接种和扩增,感染后72 h收获细胞上清液,检测病毒血凝素效价,确定病毒最佳扩增条件。随后,在搅拌瓶中进行MDCK细胞微载体悬浮培养,研究不同起始细胞接种密度（1.5×10⁵、2.0×10⁵、3.0×10⁵ 个/ml）和微载体浓度（3、5、10 g/L）对MDCK细胞生长的影响,确定细胞最佳扩增条件。根据确定的最佳扩增条件,在搅拌瓶培养体系中扩增4种病毒。结果 6孔板中4种流感病毒的最佳接种条件分别是,A/Michigan/45/2015(H1N1) pdm09：MOI 0.010 0、TPCK-胰酶浓度2 μg/ml;A/Hongkong/4801/2014(H3N2)：MOI 0.010 0、TPCK-胰酶浓度4 μg/ml;B/Brisbane/60/2008：MOI 0.001 0、TPCK-胰酶浓度4 μg/ml;B/Phuket/3073/2013：MOI 0.010 0、TPCK-胰酶浓度4 μg/ml。在搅拌瓶中以3.0×10⁵ 个/ml初始细胞密度接种,3 g/L微载体浓度培养,MDCK细胞能够实现较好的扩增,最高密度可达2.1×10⁶ 个/ml;搅拌瓶悬浮培养,以最佳接种条件接种后,4种病毒的血凝素效价为：6.75~8.42log₂ 血凝素单位/50 μl。结论   通过摸索病毒接种最佳MOI、TPCK-胰酶浓度及优化MDCK细胞微载体悬浮培养条件,能够在MDCK细胞微载体悬浮培养体系中有效扩增4价流感疫苗用病毒株,为后期工艺放大研究奠定基础。     

The action of trelibet, a new antidepressive agent on [H]noradrenaline release from rabbit pulmonary artery

Trelibet, a new antidepressant, used at 10⁻⁷–10⁻⁴ M failed to affect the [³H]noradrenaline ([³H]NA) release evoked from the isolated main pulmonary artery of the rabbit low frequency (2 Hz) nerve stimulation whether the neuronal uptake inhibitor cocaine (3 × 10⁻⁵ M) was present or not. Its metabolite (EGYT-2760) however, potentiated the nerve-evoked release of [³H]NA. In the absence of cocaine both the resting and the stimulation-evoked release of ³H increased in response to EGYT-2760. These effect were accompanied by muscle contraction. The EGYT-2760-potentiated transmitter release was inhibited either by exogenously applied 1-noradrenaline (10⁻⁶ M) or clonidine (10⁻⁶ M), preferential agonists of presynaptic ₂-adrenoceptors. The 1-noradrenaline-induced inhibition of transmitter release potentiated by EGYT-2760 was antagonized by 3 × 10⁻⁷ M yohimbine, a preferential ₂-adrenoceptor inhibitor. In the absence of cocaine, Ca²⁺ removal from the external medium failed to affect the ³H outflow-increasing effect of EGYT-2760 but abolished the nerve-evoked release-potentiating action of this compound. It is concluded that the metabolite of trelibet exerts a ‘yohimbine-like’ action, as well as a ‘tyramine-like’ effect in peripheral sympathetic nerve fibres.     

丹芍化纤胶囊对肺纤维化大鼠TGF-β1及Smad3/7表达的影响

目的 观察丹芍化纤对博来霉素致肺纤维化大鼠肺组织中TGF -β1及Smad3/7表达的影响.方法 将30只SD大鼠,随机均分为对照组、模型组和治疗组;给予模型组和治疗组大鼠的气管内注射博来霉素(5 mg·kg-1)诱导肺纤维化,对照组的气管内注射等量生理盐水;次日起ig给予治疗组大鼠丹芍化纤昆悬液(0.8g·kg-1·d-1),其余两组ig给予等量生理盐水;28 d后处死,观察各组大鼠肺组织病理变化;运用免疫组化法观察肺组织中TGF -β1及Smad3/7表达的变化.结果 模型组大鼠的肺系数明显增加,肺组织中胶原沉积明显,羟脯氨酸含量、TGF -β1和Smad3的表达增加,而Smad7表达下降;与模型组比较,治疗组大鼠的肺系数明显降低,肺组织中胶原沉积有所减轻,羟脯氨酸的含量、TGF -β1和Smad3蛋白的表达减少,而Smad7表达升高.结论 丹芍化纤具有较好的抗大鼠肺纤维化作用,其机制可能与丹芍化纤影响TGF -β1/Smads信号转导通路、下调TGF -β1及Smad3的表达,同时上调Smad7的表达有关.