内皮素受体B基因多态性与慢性阻塞性肺疾病及吸烟因素的相关性研究

目的探讨内皮素受体B（EDNRB）基因的第4外显子-30G／A处单核苷酸多态性与慢性阻塞性肺疾病（COPD）易感性及吸烟因素之间关系。方法采用PCR—RFLP技术,检测并分析EDNRB基因一30G／A（L277L）单核苷酸多态性位点在COPD组和健康对照组中的基因型频率、等位基因频率,同时分析该多态性位点与吸烟相关COPD之间是否存在相关性。结果研究发现EDNRB基因一30G／A处基因型频率分布在COPD组和对照组之间差异有统计学意义（P〈0．05）,但等位基因频率的分布在两组之间差异无统计学意义（P〉0．05）。在对COPD组和对照组中吸烟者的基因型分布频率和等位基因分布频率比较后,发现两组之间差异无统计学意义（P〉0．05）,提示该突变位点与吸烟所致的COPD之间可能无相关性存在。结论EDNRB一30G／A位点多态性可能与COPD易感性相关,与吸烟相关的COPD无关。     

血红素加氧酶-1基因多态性与冠心病发病的关系

目的：探讨血红素加氧酶-1（HO-1）基因启动子区域的GT重复序列（GTn）决定的等位基因多态性与冠心病（CHD）发病风险之间的关系。方法：采用聚合酶链式反应（PCR）扩增技术和聚丙烯酰胺凝胶电泳方法分析57例CHD患者和44例健康体检者（健康对照组）HO-1基因启动子区域GTn的分布是否存在差异。结果：与健康对照组比较,CHD组中H0—1基因启动子区S型等位基因（56．8％比38．6％）及sS基因型频率（36．4％比15．8％）显著降低,L型等位基因（43．2％比61．4％）及LL基因型频率（22．7％比38．6％）显著升高（P均〈0．05）。结论：血红素加氧酶-1基因启动子区GTn多态性可能增加冠心病的发病风险。     

DNA修复基因XRCC3多态性与肺癌易感性的关系

目的 研究DNA修复基因XRCC3多态性与肺癌易感性的关系。方法采用病例一对照研究，于2006年9月至2007年1月收集上海肺科医院原发性肺癌患者291例及同期住院的非肿瘤患者273例，应用Taqman探针结合实时荧光PCR方法分析病例组和对照组XRCC3基因Thr241Met的多态性分布，比较不同基因型与肺癌易感性的关系以及基因多态性与吸烟对肺癌的交互作用。对照组与病例组的比较用X。检验，以调整比值比及95％可信区间表示相对危险度，所有统计检验均为双侧概率检验，所有资料均用SPSS软件进行统计。结果与携带XRCC3密码子241野生纯合基因型（Thr／Thr）且不吸烟者相比，携带同样基因型且吸烟者患肺癌的风险会增加，其调整比值比（OR值）为2．47[95％可信区间（CI）为1．49-4，08，P〈0．01]；携带有杂合基因型（Thr／Met）且吸烟者患肺癌的风险也会增加，其调整OR值为2．28（95％CI为1．21-6．60，P=0，017）。野生纯合基因型（Thr／Thr）且吸烟量少于30包年可能对肺腺癌有较弱的保护作用（OR=0．49，95％CI为0．26-0．93，P=0，03）。携带有XRCC3密码子241Met等位基因且吸烟者患肺鳞癌的风险明显增加（调整OR=9．69，95％CI为3．27-28．72，P〈0．01）。携带241Met等位基因且吸烟剂量〈30包年和≥30包年的患者患肺鳞癌的风险也不同，OR值分别为8，00（95％CI为1．97-32．52，P〈0．01）和11．67（95％CI为2．98～45．73，P〈0．01）。结论DNA修复基因XRCC3多态性可能对肺癌易感性产生影响，并可能与吸烟有一定的协同作用。     

肿瘤坏死因子α-308基因多态性与慢性阻塞性肺疾病易感性的荟萃分析

目的运用荟萃分析方法综合评价肿瘤坏死因子α基因启动子308位（TNF-α-308）G／A基因型与慢性阻塞性肺疾病（COPD）的相关性。方法检索Medline和中国生物医学光盘数据库，获取TNF-α-308基因多态性与COPD易感性的病例一对照研究，使用统一的表格提取资料，应用RevMan 4．2软件进行统计学处理。结果检索的17篇文献中有18个病例-对照研究，共有1606例COPD患者（亚洲人666例，白种人940例）和2551例对照（亚洲人898例，白种人1653例）被纳入荟萃分析。亚洲人群等位基因TNF2与COPD密切相关（OR=2．62，95％CI为2．00～3．43），基因型TNF1／2及基因型TNF1／2合并TNF2／2者的COPD易感性高于基因型TNF1／1者（OR=2．44，95％CI为1．79-3．33及OR=2．78，95％CI为2．06～3．75），校正吸烟前后的敏感性结果相似。白种人等位基因TNF2与COPD易感性无相关性（OR=0．97，95％CI为0．84～1．14），基因型TNF1／2及基因型TNF1／2合并TNF2／2者的COPD易感性与基因型TNF1／1者相似（OR=0．96，95％CI为0．79～1．16及OR=1．03，95％CI为0．86～1．25），校正吸烟前后的敏感性结果相似。结论在亚洲人群中TNF2等位基因是COPD的危险因素，在白种人群中TNF-α-308的G／A基因多态性与COPD易感性无关。     

124例冠心病患者三磷酸腺苷结合盒转运体A1基因启动子区-477CT单核苷酸多态性分析

目的 研究三磷酸腺苷结合盒转运体A1基因启动子区-477C/T单核苷酸多态性与血浆高密度脂蛋白胆固醇和冠心病的关系。方法 用聚合酶链反应限制片长多态性检测124例冠心病患者和111例正常人的三磷酸腺苷结合盒转运体A1基因启动子区-477位点基因型，并比较基因型在冠心痛组与正常人组间、冠心痛组中不同临床表现型之间分布的差异性及三种基因型与冠心痛相关临床指标的关系。结果 TT基因型及T等位基因在冠心病组中的分布频率明显高于正常人组（P〈0．05和P〈0．01）。急性冠状动脉综合征组，TT基因型及T等位基因明显高于稳定型心绞痛组（P〈0．05和P〈0．01）。多支病变组TT基因型明显高于单支病变组（P〈0．05）。在冠心病组中，TT基因型血浆高密度脂蛋白胆固醇水平明显低于CC基因型（P〈0．001）。结论 三磷酸腺苷结合盒转运体A1基因启动子区-477C／T单核苷酸多态性可显著影响中国冠心痛患者血浆高密度脂蛋白胆固醇水平，而且与冠状动脉病变程度及冠心病严重程度相关。     

CD14基因-159C／T位点多态性对哮喘患者血浆IL-5的影响

目的探讨CD14基因-159C／T位点多态性和支气管哮喘（哮喘）的相关性及其对血浆IL-5的影响。方法选择正常对照组和哮喘组各150例,取外周血离心后,用酶联免疫吸附试验法测定血浆IL-5,用限制性片段长度多态性-聚合酶链反应方法检测CD14基因-159C／T位点多态性分布。结果对照组和哮喘组等位基因C、T分布有统计学差异（Х^2=10．82,P〈0．01）,C等位基因与哮喘相关（C／T的OR=1．73,95％CI=1．25—2．39,P〈0．01）,两组基因型（TT、CT、CO）频率分布有统计学差异（r=9．73,P〈0．01）;同组内C等位基因携带者血浆IL-5高于非携带者,以CC基因型最高;同一基因型哮喘组IL-5高于对照组。结论CD14基因启动子-159位点多态性与哮喘相关,C等位基因与血浆IL-5升高相关。     

白细胞介素-6基因多态性与慢性阻塞性肺疾病及吸烟因素的相关性研究

目的 探讨白细胞介素（IL）-6启动子区域基因-572C/G单核苷酸多态性与慢性阻塞性肺疾病（COPD）易感性及吸烟因素之间的关系.方法 应用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）方法,检测并分析COPD患者和健康对照者IL-6-572C/G位点基因型分布情况,同时分析该多态性位点与吸烟相关COPD之间是否存在相关性.结果 中国上海地区汉族人群存在IL-6基因-572C/G多态性;COPD组CG、GG基因型、G等位基因频率均高于对照组（P＜0.01）;基因型频率的相对风险分析发现,CG、GG基因型各自患COPD的风险分别是CC基因型的2.09倍（95％ CI：1.03 ～4.23）和5.44倍（95％ CI：1.20 ～ 24.75）;-572C/G多态性在轻中度、重度COPD组间的分布比较差异无统计学意义;该突变位点与吸烟所致的COPD之间无相关性存在.结论 IL-6基因-572G等位基因可能是中国汉族人COPD发生的易感因子,可能与携带该等位基因的人群存在IL-6水平高表达有关,其基因分布与吸烟相关的COPD无明显相关.     

结缔组织生长因子在吸烟者及慢性阻塞性肺疾病患者肺血管中的表达及其与肺血管重塑的关系

目的探讨吸烟者及慢性阻塞性肺疾病（COPD）患者的肺血管重塑中结缔组织生长因子（CTGF）的表达及意义。方法取24份（非吸烟对照组、吸烟组、吸烟伴COPD组,每组8例）手术切除的肺组织,HE染色观察肺血管重塑,天狼猩红染色检测胶原增殖,免疫组化观察CTGF在肺动脉的表达,RT-PCR检测肺动脉CTGF mRNA表达。结果（1）肺动脉管壁面积／管总面积（WA％）,非吸烟对照组为（28．4±4．7）％,吸烟组为（46．3±3．5）％,吸烟伴COPD组为（55．5±3．9）％（P〈0．01）。HE染色可见,吸烟组、吸烟伴COPD组肺动脉管壁B月显增厚。（2）肺动脉壁胶原厚度,非吸烟对照组为（6．4±1．6）μm,吸烟组为（15．9±2．4）μm,吸烟伴COPD组为（16．4±2．3）μm（P〈0．01）。天狼猩红染色可见,吸烟组、吸烟伴COPD组肺动脉管壁胶原明显增多。（3）CTGF mRNA表达量,非吸烟对照组为0．095±0．015,吸烟组为0．396±0．167,吸烟伴COPD组为0．501±0．177（P〈0．01）。（4）CTGF蛋白表达量,非吸烟对照组为0．085±0．011,吸烟组为0．245±0．095,吸烟伴COPD组为0．303±0．191（P〈0．01）。（5）相关分析：CTGF mRNA及蛋白表达量与WA％呈正相关（r分别为0．915、0．919,P〈0．01）。结论单纯吸烟者即有肺血管重塑,吸烟伴COPD者的肺血管重塑程度更重,CTGF可能在此过程中起重要作用。     

沈阳地区汉族人群SP—D基因Metl1Thr多态性与COPD易感性的关系

目的检测慢性阻塞性肺疾病（COPD）患者肺泡表面活性物质相关蛋白D（SP—D）基因MetllThr（rs721917T／C）多态性,并分析其与COPD遗传易感性的关系。方法选择100例COPD患者（COPD组）和100例年龄、性别、吸烟史相匹配的健康人（对照组）,采用PCR限制性片段长度多态性（PCR-RELP）方法检测两组SP—D基因MetllThr（rs721917T／C）位点等位基因及基因型频率分布。结果两组不同位点多态性基因型频率分布均符合遗传性Hardy—Weinberg平衡（P均〉0．05）。COPD组rITr、Tc和CC基因型频率分别为15％、46％和39％,对照组分别为25％、48％和27％,两组比较,P=0．030;COPD组C、T等位基因频率分别为62％、38％,对照组分别为51％、49％,两组比较有显著差异（P=0．026）。结论sP．D基因MetllThr多态性可能与COPD遗传易感性增加有关,C等位基因突变可能是COPD发生的重要危险因素。     

MMP-7基因-181A／G多态性与COPD易感性的相关性研究

目的探讨基质金属蛋白酶7（MMP-7）基因-18lA／G多态性与慢性阻塞性肺疾病（COPD）遗传易感性的关系。方法采用PCR-RELP方法检测100例COPD患者（COPD组）和年龄性别相匹配的100例健康对照者（健康组）MMP-7基因-181A／G多态性等位基因及基因型频率分布情况。结果两组MMP-7基因-181A／G不同位点多态性基因型频率分布均符合遗传性Hardy-Weinberg平街（P均〉0．05）。多态性检测结果显示,COPD组AA、AG和GG基因型频率分别为17％、48％和35％,健康组分别为18％,70％和12％,两组不同基因型频率分布有统计学意义（P=0．05）;COPD组G等位基因频率显著健康组（P=0．006）。结论MMP-7基因-180A／G多态性可能与COPD遗传易感性增加有关,G等位基因可能是COPD的易感基因。     

内皮素-1在大鼠肝肺综合征发病机制中的作用

目的 探讨内皮素-1(ET-1)在大鼠肝肺综合征(HPS)发病机制中的作用.方法 应用放免法检测HPS大鼠血浆和肝、肺组织匀浆中ET-1的水平.结果 ①HPS大鼠血浆和肝组织、肺组织匀浆中ET-1水平动态升高.②各阶段血浆和肝、肺组织匀浆中ET-1水平与谷丙转氨酶(ALT)、总胆红素(TBIL)呈正相关.结论 在HPS形成过程中,血浆和肝、肺组织匀浆中ET-1水平持续升高,与肝功能损害有关,提示ET-1可能参与HPS的发生.肺组织匀浆中升高的ET-1可能更多地通过与在肺血管表达增强的内皮素受体B(ETRB)结合从而扩张肺血管.     

促血管生成素-1对糖尿病大鼠肾脏色素上皮衍生因子和血小板反应蛋白-1基因表达的影响

目的 观察色素上皮衍生因子（PEDF）、血小板反应蛋白-1（TSP-1）在糖尿病大鼠肾脏的表达变化,探讨促血管生成素-1（Ang-1）对上述因子的影响. 方法 将雄性SD大鼠分正常对照（NC）组、DN组、空载处理（BV）组、Ang-1处理（AV）组.采用STZ腹腔注射诱导大鼠DN模型.成模8周后尾静脉注射Ang-1腺病毒载体.多时点检测24 hUAlb和肾组织PEDF、TSP-1蛋白及mRNA表达水平.结果 DN、BV、AV组24 hUAlb均升高,其中AV组于20周后降低（P＜0.05）.DN、BV、AV组肾组织PEDF蛋白及mRNA表达下调,TSP-1表达上调（P＜0.05）,其中AV组12周后肾组织PEDF和TSP-1mRNA及蛋白表达变化较DN、BV组改善明显（P＜0.05）. 结论 糖尿病大鼠肾脏PEDF、TSP-1异常表达参与DN发生发展,给予Ang-1可改善糖尿病大鼠肾脏PEDF、TSP-1异常表达.     

肾上腺髓质素和内皮素-1在大鼠肝肺综合征发病机制中的作用

[目的]探讨肝肺综合征(HPS)的发病机制.[方法]采用胆总管结扎(CBDL)术制备大鼠HPS模型,观察肺组织肾上腺髓质素(ADM)、内皮素-1(ET-1)及其受体(ETRA和ETRB)的表达和分布.[结果]在大鼠HPS形成过程中,血浆和肺组织中ADM、ET-1水平动态升高,且与肺泡-动脉氧分压差(A-aDO2)正相关;HPS大鼠肺组织中ADM、内皮素前体原(ppET-1 mRNA)的表达较假手术组明显增强,差异均有统计学意义(P＜0.05).HPS大鼠肺血管ETRA的分布及染色强度与假手术组比较无明显变化,而ETRB在远端肺小动脉和小静脉内膜上表达明显增强.图像分析结果显示CBDL 5周(w)组大鼠ETRA染色面积、平均积分光密度值与假手术组比较差异无统计学意义(P＞0.05),而CBDL 5 w组大鼠ETRB染色面积和平均积分光密度值明显高于假手术组,差异均有统计学意义(P＜0.05).[结论]扩血管物质ADM和缩血管物质ET-1的共同作用可能参与HPS的发生,肺组织中升高的ET-1可能更多地通过与在肺血管表达增强的ETRB结合从而扩张肺血管.     

Evaluation of the efficacy and safety of anti-PD-1 and anti-PD-L1 antibody in the treatment of non-small cell lung cancer (NSCLC): a meta-analysis

Background

Currently, blockade of the programmed cell death 1 (PD-1)/PD-1 ligand 1 (PD-L1) signaling pathway has been proved one of the most promising immunotherapeutic strategies against cancer. Several antibodies have been developed to either block the PD-1 or its ligand PD-L1 are under development. So far, a series of phase I trials on PD-1/PD-L1 antibodies for non-small cell lung cancer (NSCLC) have been completed, without reports of results from phase II studies. Thus, we sought to perform a meta-analysis incorporating all available evidences to evaluate the efficacy and safety of PD-1 or PD-L1 inhibition therapy.

Methods

Electronic databases were searched for eligible literatures. Data of objective respond rate (ORR) and rate of adverse effects (AEs) with 95% confidence interval (CI) evaluated by immunohistochemistry (IHC) was extracted. The outcomes were synthesized based on random-effect model. Subgroup analyses were proposed.

Results

In overall, ORR in the whole population with PD-1 blockage treatment is 22.5% (95% CI: 17.6% to 28.2%). Additionally, the rate of Grade 3-4 AEs is 16.7% (95% CI: 6.5% to 36.8%) and drug-related death rate is 2.5% (95% CI: 1.3% to 4.6%). As for patients with PD-L1 inhibition therapy, an overall ORR is 19.5% (95% CI: 13.2% to 27.7%). A higher rate of Grade 3-4 AEs (31.7%, 95% CI: 14.2% to 56.5%) is observed with a lower drug-related death rate (1.8%, 95% CI: 0.4% to 8.3%). In exploratory analyses of anti-PD-1 agents, we observed that greater ORR was presented in the median-dose cohort (3 mg/kg) than that of both low-dose (1 mg/kg) and high-dose (10 mg/kg) cohort (low-dose vs. median-dose: OR =0.12, P=0.0002; median-dose vs. high-dose: OR =1.47, P=0.18).

Conclusions

Anti-PD-1 and anti PD-L1 antibodies showed objective responses in approximately one fourth NSCLC patients with a tolerable adverse-effect profile. In addition, median-dose (3 mg/kg) might be a preferential dosage of anti-PD-1 agents.     

The role of interleukin-1 receptor antagonist in the prevention and treatment of disease

Abstract

?Interleukin-1 (IL-1) and tumor necrosis factor α (TNF-α) play key proinflammatory roles in a variety of human diseases, including rheumatoid arthritis (RA). IL-1 receptor antagonist (IL-1Ra) is a naturally occurring structural variant of IL-1 that competitively inhibits receptor binding of IL-1. Four forms of IL-1Ra have been described: secretory IL-1Ra (sIL-1Ra) and three intracellular molecules (icIL-1Ra1, 2, and 3). Excess amounts of IL-1Ra are necessary to inhibit the biological effects of IL-1. The endogenous production of IL-1Ra plays an anti-inflammatory role, but the level of production of IL-1Ra in inflamed tissues may not be adequate to block IL-1 effectively. An allelic polymorphism in the IL-1Ra gene is associated with a variety of human diseases, largely of epithelial or endothelial cell origin. The disease associated allele IL1RN*2 may lead to a decreased production of icIL-1Ra1 by these cells, predisposing the patient to an imbalance in the IL-1 system. The therapeutic administration of IL-1Ra was found to be safe and efficacious in the treatment of RA. Intraarticular delivery of the IL-1Ra cDNA by ex vivo gene therapy in patients with RA was effective in enhancing local IL-1Ra production. This unique form of therapy is under further evaluation.     

Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel

The study of hematopoietic colony-forming units using semisolid culture media has greatly advanced the knowledge of hematopoiesis. Here we report that similar methods can be used to study pancreatic colony-forming units. We have developed two pancreatic colony assays that enable quantitative and functional analyses of progenitor-like cells isolated from dissociated adult (2–4 mo old) murine pancreas. We find that a methylcellulose-based semisolid medium containing Matrigel allows growth of duct-like “Ring/Dense” colonies from a rare (∼1%) population of total pancreatic single cells. With the addition of roof plate-specific spondin 1, a wingless-int agonist, Ring/Dense colony-forming cells can be expanded more than 100,000-fold when serially dissociated and replated in the presence of Matrigel. When cells grown in Matrigel are then transferred to a Matrigel-free semisolid medium with a unique laminin-based hydrogel, some cells grow and differentiate into another type of colony, which we name “Endocrine/Acinar.” These Endocrine/Acinar colonies are comprised mostly of endocrine- and acinar-like cells, as ascertained by RNA expression analysis, immunohistochemistry, and electron microscopy. Most Endocrine/Acinar colonies contain beta-like cells that secrete insulin/C-peptide in response to D-glucose and theophylline. These results demonstrate robust self-renewal and differentiation of adult Ring/Dense colony-forming units in vitro and suggest an approach to producing beta-like cells for cell replacement of type 1 diabetes. The methods described, which include microfluidic expression analysis of single cells and colonies, should also advance study of pancreas development and pancreatic progenitor cells.     

包虫病患者血清中sICAM-1水平的检测

目的通过检测包虫病患者血清sICAM-1水平,为探讨包虫免疫逃避机理提供资料。方法以ELISA双抗体夹心法检测40名包虫病患者和16名健康体检者血清中sICAM-1水平。结果包虫病患者和健康体检者血清中sICAM-1的水平分别为(106.76±15.68)pmol/L和(43.42±31.41)pmol/L,两者间差异有显著性(P<0.05)。结论人体感染包虫后,sICAM-1的合成增加。     

The role of interleukin-1 receptor antagonist in the prevention and treatment of disease

 Interleukin-1 (IL-1) and tumor necrosis factor α (TNF-α) play key proinflammatory roles in a variety of human diseases, including rheumatoid arthritis (RA). IL-1 receptor antagonist (IL-1Ra) is a naturally occurring structural variant of IL-1 that competitively inhibits receptor binding of IL-1. Four forms of IL-1Ra have been described: secretory IL-1Ra (sIL-1Ra) and three intracellular molecules (icIL-1Ra1, 2, and 3). Excess amounts of IL-1Ra are necessary to inhibit the biological effects of IL-1. The endogenous production of IL-1Ra plays an anti-inflammatory role, but the level of production of IL-1Ra in inflamed tissues may not be adequate to block IL-1 effectively. An allelic polymorphism in the IL-1Ra gene is associated with a variety of human diseases, largely of epithelial or endothelial cell origin. The disease associated allele IL1RN^*2 may lead to a decreased production of icIL-1Ra1 by these cells, predisposing the patient to an imbalance in the IL-1 system. The therapeutic administration of IL-1Ra was found to be safe and efficacious in the treatment of RA. Intraarticular delivery of the IL-1Ra cDNA by ex vivo gene therapy in patients with RA was effective in enhancing local IL-1Ra production. This unique form of therapy is under further evaluation. Correspondence to:W.P. Arend     

Sequence heterogeneity of the merozoite surface protein-1 gene (MSP-1) of Plasmodium vivax wild isolates in southeastern Iran

The merozoite surface protein of Plasmodium vivax (PvMSP-1) has been considered as a vaccine candidate, which exhibits antigenic diversity among isolates. We investigated the extent of sequence variation in the polymorphic region 5 of PvMSP-1 in order to characterize the genetic structure and composition of P. vivax in clinical isolates from Iranshahr and Chahbahar districts of Sistan and Baluchistan province, Iran. The PvMSP-1 gene amplification revealed size variation among the isolates, ranging from 430 to 550 bp. Sequences were obtained for 15 Iranian and one Pakistani isolates and 14 different alleles were detected. Results also showed three distinct sequence types of the polymorphic region. Sequence analysis has shown several single nucleotide polymorphisms to occur in this block of PvMSP-1, creating different alleles in the progeny and also microheterogeneity in the region. Thus, this study provides preliminary evidence of sequence heterogeneity in the Iranian P. vivax population.     

软脉胶囊对血管性痴呆大鼠脑组织ET-1、CGRP及海马CA1区n-NOS表达的影响

目的探讨软脉胶囊治疗血管性痴呆的机制。方法采用双侧颈总动脉结扎，制作慢性脑灌注不足动物模型，观察软脉胶囊对大鼠脑组织ET-1、CGRP及海马CA1区n-NOS表达的影响。结果软脉胶囊可降低大鼠脑组织中ET-1，升高大鼠脑组织中CGRP。同时可明显提高大鼠海马CA1区n-NOS的表达。结论软脉胶囊对慢性脑灌注不足大鼠脑组织具有保护作用。