HLA-DRB和ACE基因与肺结核的相关性分析

目的探讨人类白细胞抗原(HLA)-DRB基因和血管紧张素转换酶(ACE)基因多态性与肺结核的相关性。方法采用聚合酶链反应-序列特异性引物法(PCR-SSP)对肺结核组和健康对照组进行HLA-DRB和ACE基因分型。结果肺结核组HLA-DRB₁^*15等位基因的频率显著高于对照组(P＜0.05);复治和耐药肺结核组ACE基因的I/D基因型频率显著高于初治和无耐药组(P＜0.05)。结论HLA-DRB₁^*15等位基因及ACE基因的I/D基因型与肺结核密切相关。     

蜡样芽胞杆菌在不同标本中的毒力基因分布特征

目的 研究蜡样芽胞杆菌毒力基因在不同标本中的分布特征。方法 分离自环境监测标本(米粉、奶粉、土壤)和疾病相关标本(米饭、凉皮、眼内炎和肿瘤患者)的蜡样杆菌333株,PCR扩增蜡样杆菌11个毒力基因,包括溶血性BL基因(hblC、hblD、hblA、hblB)、非溶血性基因(nheA、nheB、nheC)、肠毒素FM基因和T基因(entFM、bceT)、细胞毒素K基因(cytK)和呕吐毒素相关基因(ces),统计不同标本菌株中毒力基因的携带数目和各毒力基因的携带率,方差分析和卡方检验比较毒力基因在不同标本中,尤其是在疾病相关标本与环境监测标本中的携带差异。结果 研究菌株携带毒力基因平均数目为5.97个,88.29%的菌株携带至少3个毒力基因,12.31%的菌株携带除ces外的所有基因。标本携带毒力基因数目从高到低依次为患者(8.75)、凉皮(8.20)、米饭(7.13)、土壤(6.22)、米粉(5.78)和奶粉(5.71)。患者、凉皮分别与奶粉的基因数目两两比较有统计学差异显著性。菌株各毒力基因的携带率从高到低依次为非溶血性基因(89.19%)、entFM基因(79.88%)、bceT基因(49.85%)、溶血性BL基因(48.35%)、ctyK基因(47.75%)和ces基因(1.50%)。溶血性基因在疾病相关标本中的携带率比环境监测标本高(χ²=8.230,P<0.01),其余基因携带率则在两类标本中无统计学差异。环境监测标本中,土壤的溶血性基因携带率高于米粉和奶粉(χ²=15.071,P<0.01),非溶血性基因和entFM基因的携带率则低于后两者,检验值分别为(χ²=9.603,P<0.05)和(χ²=21.634,P<0.01)。结论 蜡样杆菌毒力基因在不同标本中的分布,尤其是毒力基因数目和溶血性基因在疾病相关标本中较高的携带特点,为研究蜡样杆菌的致病性提供了一定的参考依据,具有一定的临床意义。     

IL-1α基因启动子-889C/T和IL-1Ra第2内含子VNTR基因型及血清水平与2型糖尿病的相关性研究

目的　研究白细胞介素 1α(IL 1α)基因启动子 889C/T多态性和白细胞介素 1受体拮抗剂 (IL 1Ra)第 2内含子可变数目串联重复(VNTR)多态性及血清水平与 2型糖尿病 (T2DM)的相关关系。方法　采用聚合酶链反应 限制性片段长度多态性 (PCR RFLP)技术 ,检测 1 35例T2DM患者和 1 4 0例健康对照者的IL 1α( 889C/T)和IL 1Ra(VNTR)基因型 ,同时采用ELISA检测T2DM患者和对照者的血清IL 1水平。结果　T2DM组血清IL 1水平显著高于对照组 (P <0 0 1 ) ,IL 1α基因 889C/T基因型频率和等位基因频率在T2DM组和对照组比较差异有显著性 (P <0 0 5) ,等位基因频率的相对风险分析发现 ,T等位基因携带者患T2DM的风险是C等位基因的 2 0 2 3倍 (OR =2 0 2 3 ,95 %CI:1 1 4 7～ 3 567) ;IL 1Ra基因型频率和等位基因频率在T2DM组和对照组比较差异有显著性 (P <0 0 5) ,与健康对照组比较 ,不携带IL 1RaⅡ的基因型患 2型DM的相对风险度增加 2 0 2 5倍 (OR =2 0 2 5 ,95 %CI :1 0 33～ 3 967) ,携带IL 1RaⅡ的基因型的T2DM患者血清IL 1Ra水平显著高于不携带者〔(1 4 2 6 2 5± 32 4 31 ) pg/mlVS (1 2 99 56± 2 93 47) pg/ml,P <0 0 1〕。 结论　IL 1α( 889C/T)和IL 1Ra(VNTR)基因多态性与T2DM的发病具有相关性 ,其     

重组双基因鸭源大肠杆菌菌蜕的制备

目的 利用裂解E基因和葡萄球菌核酸酶A(SN)基因的表达,制备鸭源大肠杆菌(E.coli)菌蜕。方法 通过将带有裂解 E基因和葡萄球菌核酸酶A(SN)基因的质粒 pET29a-E-S 转化至鸭源E.coli O₉₂中,对E.coli O₉₂(pET29a-E-S)进行诱导,每隔 30 min 检测菌液的OD₆₀₀值,并检测培养液中所含 DNA。结果 通过诱导,E.coli O₉₂(pET29a-E-S) OD₆₀₀值在诱导 90 min 后开始持续下降,180 min 时开始趋于平稳,到 360 min 溶菌效率达 99.999%。并且葡萄球菌核酸酶把 DNA 降解为 50 ~400 bp 的小片断。结论 通过裂解 E基因和 SN 基因表达,成功制备了E.coli O₉₂菌蜕,本实验为进一步研究该菌蜕疫苗奠定了基础。     

成都地区肉鸭屠宰链弯曲菌毒力基因分布及分子分型研究

目的 了解并分析肉鸭屠宰环节中弯曲菌分离株的9种毒力基因分布及分子分型特征。方法 利用特异性引物对弯曲菌9种与致病力相关的毒力基因进行PCR检测;参照美国PulseNet 脉冲场凝胶电泳(PFGE)标准方法,对68株鸭源弯曲菌分离株进行PFGE分型。结果 毒力基因PCR检测结果显示空肠弯曲菌中cadF、iamA和cheY毒力基因携带率均为100%,flaA(97.1%)、cdtB(94.3%)、cdtC(94.3%)和ciaB(80%)毒力基因的携带率也较高,其余毒力基因cdtA(25.7%),virB11(2.9%)较低;结肠弯曲菌中除cadF(100%)毒力基因外,高于50%携带率的毒力基因有cheY(84.8%)、cdtB(72.7%)、iamA(66.7%)和cdtA(54.5%),另外4个毒力基因flaA(48.5%)、virB11(18.2%)、cdtC(36.4%)和ciaB(18.2%)携带率均较低。利用PFGE方法对弯曲菌分离株进行分子分型,结果显示35株空肠弯曲菌和33株结肠弯曲菌可分为15个和11个谱型,表现为较低的遗传多样性。结论 弯曲菌毒力基因分布广泛,且空肠弯曲菌携带率较高;PFGE基因谱型相似性表明该肉鸭屠宰场存在交叉感染和弯曲菌沿屠宰链传播的现象。     

310例中国人基因组干扰素α1/α2基因多态性研究

目的 若药品α-干扰素亚型与人体体内基因型不一致,接受抗病毒治疗的患者可被诱导产生更多的抗干扰素中和抗体,导致不应答和治疗失败。通过分析中国人群干扰素α1(IFNA1)和α2(IFNA2)基因多态性,可帮助阐明重组人干扰素α1和α2在中国人群中的遗传背景,为临床用药提供参考。方法 在单核苷酸多态性数据库中分别检索IFNA1基因和IFNA2基因编码区错义突变单核苷酸多态性(SNP)位点,再在千人基因组数据库查询收录的中国人IFNA1和IFNA2等位基因的分布情况,分析两种基因多态性并计算出相关的等位基因频率。结果 在千人基因组数据库共收录310例中国人双倍染色体基因组序列,所有中国人均有IFNA1和IFNA2基因座,其中,IFNA1基因座含有IFNα1、IFNα1(Ala137Val)和IFNα1(Arg148Gln) 三种等位基因,基因频率分别为99.6%、0.2%和0.2%;IFNA2基因座也含有IFNα2b、IFNα2b(Leu140Val)和IFNα2b(Ala120Thr)三种等位基因,基因频率分别为98.4%、0.2%和1.4%。结论 IFNα1和IFNα2b分别是中国人群IFNA1和IFNA2基因座的优势等位基因,在我国批准上市的IFNα1b、IFNα2a和IFNα2b三种亚型α-干扰素中,仅有IFNα2b与中国人基因型一致。     

熔解曲线分析法对济南地区汉族人群HLA-A、B和HPA1-18,21基因多态性分布的研究

目的:探讨济南地区汉族人群血小板抗原(HPA)1-18,21和人类白细胞抗原(HLA-A、B)的基因多态性分布,为建立本地区机采血小板献血者基因分型资料库奠定基础。方法:采用荧光PCR熔解曲线分析法对本站随机选取的347例无亲缘关系固定献血者进行HLA-A、B和HPA1-18,21基因分型。结果:HLA-A、B位点分别检测出18个和34个等位基因,基因频率较高的分布为A^*02(0.278 1),A^*11(0.168 6),A^*24(0.147 0)与B^*13(0.142 7)、B^*60(0.069 2)、B^*61(0.064 8)、B^*62(0.063 4)。最常见的HPA基因组合型为：HPA-1a/1a-2a/2a-3a/3b-4a/4a-5a/5a-6a/6a-(7-14)a/a-15a/15b-(16-18)a/a-21a/21a。结论:济南汉族人群HLA和HPA分布具有多态性,与其他地区和种族比较有差异性,应建立本地区血小板捐...     

耻垢分枝杆菌新型毒素-抗毒素系统MSMEG_3435-3436基因功能的初步研究

目的 初步探讨耻垢分枝杆菌(Mycobacterium smegmatis)毒素-抗毒素(toxin-antitoxin,TA)系统基因的功能及其在细菌药物耐受中的作用。方法 利用无水四环素(ATc)诱导穿梭质粒构建毒素基因(MSMEG_3436和MSMEG_6760)表达系统,检测毒素基因表达的抑菌作用。应用CRISPR-Cas12a基因编辑技术构建ΔMSMEG_3435-3436和ΔMSMEG_6762-6760敲除菌株,探究毒素-抗毒素系统对菌株生长的影响。通过计算菌株存活率检测MSMEG_3435-3436基因对异烟肼(96μg/ml)和利福平(40μg/ml)的耐受相关性。在耻垢分枝杆菌中用LacZ报告基因分别替换毒素-抗毒素基因(MSMEG_1277-1278、 MSMEG_1283-1284、 MSMEG_3435-3436、 MSMEG_4447-4448和MSMEG_5635-5634),构建5个启动子活性检测突变菌株(SY3328、SY3309、SY6407、SY3310和SY3311),并将pMV261空载体和pMV261-抗毒素系列质粒分别电转至5个突变菌株中,通过测定吸光度值(A₆₀₀、A₅₅₀和A₄₂₀)计算β-半乳糖苷酶活性[酶活性单位为“Miller单位(MU)”],以检测毒素抗毒素系统的启动子活性。结果 在耻垢分枝杆菌中,ATc诱导表达毒素基因MSMEG_3436可抑制细菌生长,而同时表达对应的抗毒素基因MSMEG_3435可消除抑制作用;ATc诱导表达毒素基因MSMEG_6760未发现明显的抑菌作用。与野生株相比,ΔMSMEG_3435-3436和ΔMSMEG_6762-6760敲除菌株在7H9液体培养基中生长表型无明显差异。野生株和ΔMSMEG_3435-3436敲除菌株经异烟肼和利福平处理后的存活率[分别为(4.38±1.48)%和(3.49±0.66)%,(0.15±0.04)%和(0.03±0.02)%]显示毒素-抗毒素基因MSMEG_3435-3436与药物耐受性无关(t=0.548,P=0.613;t=2.663,P=0.056)。启动子(SY3328、SY3309、SY6407、SY3310和SY3311)在携带pMV261-空载体和pMV261抗毒素表达质粒的报告菌株中的β-半乳糖苷酶活性分别为(376.50±17.13)和(315.50±20.71)、(189.00±12.24)和(160.70±9.89)、(225.20±9.95)和(211.70±2.57)、(221.40±12.07)和(186.60±13.17)、(179.10±5.87)和(127.70±19.21)MU,差异均无统计学意义(t=2.272,P=0.086;t=1.795,P=0.147;t=1.319,P=0.258;t=1.949,P=0.123;t=2.562,P=0.063)。结论 成功构建了MSMEG_3435-3436和MSMEG_6762-6760在耻垢分枝杆菌中的诱导表达体系及敲除菌株,并发现MSMEG_3435-3436是一个新的有功能的毒素-抗毒素系统,以及这两个毒素-抗毒素系统与菌株生长表型及异烟肼和利福平耐受性无关,最后发现耻垢分枝杆菌中5对毒素-抗毒素系统的抗毒素基因可能在自身启动子调控中不发挥关键作用,可为进一步研究结核分枝杆菌毒素-抗毒素系统的功能提供线索。     

皖南地区HLA-Brs34933313基因多态性与肾综合征出血热的相关性分析

目的 探讨皖南地区汉族人群HLA-B基因单核苷酸多态性(Single Nucleotide Polymorphisms,SNP)rs34933313与肾综合征出血热(Hemorrhagic fever with renal syndrome,HFRS)的相关性。方法 采集HFRS患者14例(病例组)和健康组50例(对照组)新鲜血样,提取基因组DNA,通过等位基因特异引物-聚合酶链反应(ASP-PCR)对HLA-B基因位点分型,应用SPSS22.0统计分析其等位基因及基因型两组间的差异。结果 HLA-B基因rs34933313位点受试人群分布符合哈迪-温伯格(Hardy-Weinberg,H-W)遗传平衡,两组等位基因G、C(χ²=4.38,OR=2.45,P=0.04)和基因型GG、GC、CC(χ²=6.47,P=0.04)频率分布差异均有统计学意义;病例组中C等位基因的分布频率高于G等位基因,且携带C等位基因罹患HFRS的概率是G等位基因的2.45倍;亚组分析发现 HLA-B基因rs34933313位点在显性遗传模型下(χ²=6.45,P=0.01,OR=10.21,95%CI=1.24~84.18)与HFRS发病风险具有相关。结论 皖南地区汉族人群HLA-B基因rs34933313位点与HFRS发病风险存在相关性,且携带C等位基因者会提高HFRS的发生风险。     

大肠杆菌肠毒素基因多重PCR检测方法的建立

目的 建立一种快速检测大肠杆菌耐热肠毒素(heat-stable enterotoxin, STa, STb)和不耐热肠毒素(heat-labile enterotoxin, LT-Ⅰ, LT-Ⅱ)基因的多重PCR方法。方法 参照文献合成四对可扩增产肠毒素大肠杆菌(Enterotoxigenic Escherichia coli, ETEC)耐热肠毒素基因(estA、estB)和不耐热肠毒素基因(elt-Ⅰ、elt-Ⅱ)的特异性引物,通过反应条件的优化,敏感性、特异性试验和临床样品检测,建立检测大肠杆菌肠毒素的多重PCR方法。结果 用所建立的多重PCR方法可特异性扩增出estA(229 bp)、estB(480 bp)、elt-Ⅰ(605 bp)和elt-Ⅱ(300 bp)基因片段,最低检出量分别为2.55×10¹ CFU/μL、2×10¹ CFU/μL、2×10¹ CFU/μL和2.47×10³ CFU/μL。从22株大肠杆菌分离株中检测到estA基因(2/22),elt-Ⅱ基因(3/22),未检测到estB和elt-Ⅰ基因,检测结果与常规PCR检测结果一致。结论 建立了检测大肠杆菌肠毒素基因(estA、estB、elt-Ⅰ和elt-Ⅱ)的多重PCR方法,该方法具有良好的特异性和敏感性,能够满足对细菌培养物的检测要求。     

125株耐多药结核分枝杆菌的吡嗪酰胺耐药及pncA基因突变分析

目的 了解125株耐多药结核分枝杆菌对吡嗪酰胺(PZA)的耐药水平和pncA基因的突变特征,为PZA耐药的快速诊断提供依据。方法 通过简单随机抽样方法选取2010—2019年福建省9个耐药监测点的125株耐多药菌株,采用BACTEC MGIT 960液体培养进行PZA药物敏感性试验(简称“药敏试验”),通过PCR扩增pncA目的基因,分析耐多药菌株对PZA的耐药性及pncA基因突变情况。结果 50株耐多药菌株对PZA耐药,PZA耐药率为40.0%(50/125)。男性患者PZA耐药率为39.0%(37/95),女性患者PZA耐药率为43.3%(13/30),差异无统计学意义(χ²=0.183,P=0.669);15~25岁、26~45岁、46~60岁、61~80岁患者PZA耐药率分别为58.3%(7/12)、 32.7%(16/49)、39.5%(17/43)、 47.6%(10/21),PZA耐药率在年龄组之间的差异无统计学意义(χ²=3.294,P=0.348);初治患者PZA耐药率为41.5%(21/65),复治患者PZA耐药率为38.3%(23/60),差异无统计学意义(χ²=0.134,P=0.715)。30株PZA耐药菌株检出pncA基因突变,突变类型22种,突变检出率为60.0%(30/50);PZA耐药株的pncA基因突变检出率[60.0%(30/50)]高于PZA敏感株的pncA基因突变检出率[5.3%(4/75)],差异有统计学意义(χ²=45.276,P=0.000)。结论 福建省耐多药结核分枝杆菌的PZA耐药率较高,且 pncA基因突变检出率较高,临床上应关注PZA的耐药问题。     

Detection of the association between a deletion polymorphism in the gene encoding angiotensin I-converting enzyme and advanced diabetic retinopathy

We investigated the relationship between advanced diabetic retinopathy (ADR) and an angiotensin-converting enzyme (ACE) gene polymorphism in subjects with type 2 diabetes and ADR, pre-proliferative (PrePDR) or proliferative diabetic retinopathy (PDR) without overt nephropathy. Polymerase chain reactions were used to detect insertion/deletion (I/D) polymorphisms of the ACE gene. There was no difference in the frequency of II, ID, or DD genotypes, or of I and D alleles among subjects with type 2 diabetes without diabetic retinopathy (NDR) or with simple diabetic retinopathy (SDR) and non-diabetic controls. There was also no difference in the frequency of ACE genotypes among subjects with type 2 diabetes with NDR, or SDR and ADR. However, the frequency of the ACE DD genotype in ADR was significantly higher than that in controls (χ²=6.64, P=0.036). On the other hand, the frequency of the D allele in ADR was significantly higher than that in controls (χ²=6.33, P=0.012), NDR (χ²=4.18, P=0.041) and SDR (χ²=4.89, P=0.027), respectively. These results indicate a significant relationship between the presence of the D allele polymorphism in the ACE gene and ADR in Japanese subjects with type 2 diabetes and no overt nephropathy.     

Room for improvement in the treatment of pancreatic cancer: Novel opportunities from gene targeted therapy

Pancreatic cancer is one of the highest and in fact, unchanged mortality-associated tumor, with an exceptionally low survival rate due to its challenging diagnostic approach. So far, its treatment is based on a combination of approaches (such as surgical resection with or rarely without chemotherapeutic agents), but with finite limits. Thus, looking for additional space to improve pancreatic tumorigenesis therapeutic approach, research has focused on gene therapy with unexpectedly growing horizons not only for the treatment of inoperable pancreatic disease, but also for its early stages. In vivo gene delivery viral vectors, despite few disadvantages (possible immunogenicity, toxicity, mutagenicity, or high cost), could be one of the most efficient cancer gene therapeutic strategies for clinical application due to their superiority compared with other systems (ex vivo delivery strategies). Their dominance consists of simple preparation, easy operation and a wide range of functions. Adenoviruses are one of the most common used vectors, inducing strong immune as well as inflammatory reactions. Oncolytic virotherapy, using the above mentioned in vivo viral vectors, is one of the most promising non-pathogenic, highly-selective cytotoxic anti-cancer therapy using anti-cancer agents with high anti-tumor potency and strong oncolytic effect. There have been a variety of targeted therapeutic and pre-clinical strategies tested for gene therapy in pancreatic cancer such as gene-editing systems (e.g., clustered regularly interspaced palindromic repeats-Cas9), RNA interference technology (e.g., microRNAs, short hairpin RNA or small interfering RNA), adoptive immunotherapy and vaccination (e.g., chimeric antigen receptor T-cell therapy) with encouraging results.     

miR-20a-5p调控结核分枝杆菌诱导巨噬细胞凋亡相关基因表达的研究

目的 明确小非编码RNA(microRNA,miR-20a-5p)对结核分枝杆菌(MTB)诱导人巨噬细胞凋亡相关基因表达的调控作用。方法 构建可表达或抑制miR-20a-5p、转染miR-20a-5p抑制剂(miR-20a-5p-inhibitor,简称“miR-20a-5p-inh”)、作为慢病毒载体的阴性对照(LV1-NC)的慢病毒载体,由oligo-dT引物通过固相亚磷酰胺法合成茎环寡核苷酸,并克隆到含绿色荧光蛋白(GFP)的慢病毒载体pGLV3-GFP内,将构建的miR-20a-5p、miR-20a-5p-inh、LV1-NC转染THP-1人巨噬细胞3h,培养72h后用流式细胞仪分选出GFP+THP-1细胞,并分别采用减毒MTB菌株(H37Ra)感染8h和过氧化氢(H₂O₂)处理30min 两种方法诱导。通过实时定量PCR技术测定细胞中线粒体相关抗凋亡基因Bcl-2,以及促凋亡基因Bax、Bim和Bad的转录水平。同时,用蛋白免疫印迹法检测细胞裂解物中相关蛋白的表达。结果 慢病毒转染细胞后,在无刺激的THP-1细胞中miR-20a-5p的相对荧光强度值为12.21±1.29、miR-20a-5p-inh为9.68±1.38、LV1-NC为10.64±0.96,三者的表达差异均无统计学意义(q=1.815,P=0.385;q=2.072,P=0.602);但在H37Ra和H₂O₂的刺激后,miR-20a-5p过表达时其相对荧光强度值分别为7.20±0.53、8.55±0.82,明显高于LV1-NC (4.46±0.07、5.49±0.44)(q=50.250,P=0.007;q=1.041,P<0.01),miR-20a-5p受抑制时(1.88±0.08、1.44±0.21)明显低于LV1-NC(q=3.457,P=0.031;q=4.384,P=0.001)。在感染miR-20a-5p慢病毒的THP-1细胞中,H37Ra诱导Bcl-2的表达增加(相对荧光强度值由10.67±0.89增至14.98±0.88)(q=1.064,P=0.008),而感染miR-20a-5p-inh慢病毒时Bcl-2表达减少(由10.67±0.89降至6.49±0.47)(q=3.518,P=0.003);在miR-20a-5p过表达的THP-1细胞中Bim的转录水平减少(由1.22±0.05降至0.98±0.04)(q=1.240,P=0.011),当miR-20a-5p受抑制时Bim的转录水平增加(由1.22±0.05增至1.51±0.08)(q=2.460,P=0.021)。蛋白印迹法检测凋亡相关基因Bcl-2和Bim的蛋白表达水平,结果与基因转录水平相符。 结论 miR-20a-5p的表达水平影响MTB诱导的巨噬细胞凋亡相关基因Bcl-2和Bim的表达,并且与促凋亡基因Bim的水平呈负相关,提示miR-20a-5p可能通过调控Bim的表达而影响细胞凋亡。     

Duodenal epithelial transport in functional dyspepsia: Role of serotonin

AIM: To investigate functional duodenal abnormalities in functional dyspepsia (FD) and the role of serotonin (5-hydroxytryptamine, 5-HT) in mucosal ion transport and signalling. METHODS: Duodenal mucosal biopsies were obtained from 15 patients with FD and 18 healthy controls. Immunohistochemistry was used to study the number of 5-HT-containing cells and real-time polymerase chain reaction for expression of 5-HT receptors 1A, 1B, 2A, 2B, 3A, 3B, 3C, 3D, 3E, 4 and 7, as well as expression of the serotonin re-uptake transporter (SERT) gene SLC6A4 and tryptophan hydroxylase 1 (TPH1). Biopsies were mounted in Ussing chambers for evaluation of basal and 5-HT-stimulated short-circuit current (SCC). RESULTS: Conductance was lower in FD [42.4 ± 4.7 mS/cm² (n = 15) vs 62.5 ± 4.5 mS/cm² (n = 18), P = 0.005]. 5-HT induced a dose dependent rise in SCC in both FD (n = 8) and controls (n = 9), the rise was lower in FD (P < 0.001). Mean number of 5-HT stained cells per high power field was the same [34.4 ± 8.4 in FD (n = 15) and 30.4 ± 3.7 in controls (n = 18), P = 0.647]. The following genes were highly expressed: 5-HT receptor HTR3E, HTR4, HTR7, SERT gene (SLC6A4) and TPH1. Differences in expression levels were observed for HTR3E (higher expression in FD, P = 0.008), HTR7 (lower expression in FD, P = 0.027), SLC6A4 (higher expression in FD, P = 0.033) and TPH1 (lower expression in FD, P = 0.031). CONCLUSION: Duodenal ion transport in response to exogenous 5-HT is abnormal in FD patients and associated with high expression of the HTR3E receptor and the serotonin transporter.     

Expression of the early-late gene encoding the nuclear receptor HR3 suggests its involvement in regulating the vitellogenic response to ecdysone in the adult mosquito

The insect steroid hormone, 20-hydroxyecdysone (20E), is a key factor controlling critical developmental events of embryogenesis, larval molting, metamorphosis, and, in some insects, reproduction. We are interested in understanding the molecular basis of the steroid hormone ecdysone action in insect egg development. The yellow fever mosquito, Aedes aegypti, in addition to being an important vector of human diseases, represents an outstanding model for studying molecular mechanisms underlying egg maturation due to stringently controlled, blood meal-activated reproductive events in this insect. To elucidate the genetic regulatory hierarchy controlling the reproductive ecdysone response, we have investigated ecdysone-regulated gene expression in vitellogenic mosquito ovaries and fat bodies. We have previously demonstrated the conservation of a primary ecdysone-triggered regulatory hierarchy, implicated in development of immature stages of Drosophila, represented by the ecdysone receptor/Ultraspiracle complex and an early gene E75 during the reproductive ecdysone response (Wang, S.-F., Miura, K., Miksicek, R.J., Segraves, W.A., Raikhel, A.S., 1998. DNA binding and transactivation characteristics of the mosquito ecdysone receptor — Ultraspiracle complex. J. Biol. Chem. 273, 27531–27540; Pierceall, W.E., Li, C., Biran, A., Miura, K., Raikhel, A.S., Segraves, W.A., 1999. E75 expression in mosquito ovary and fat body suggests reiterative use of ecdysone-regulated hierarchies in development and reproduction. Mol. Cell. Endocrinol. 150, 73–89). The present paper demonstrates that conservation of the factors involved in the ecdysone-responsive genetic hierarchy regulating female reproduction extends beyond the early genes. Here, we identify AHR3, a highly conserved homologue of the Drosophila HR3 early-late ecdysone-inducible gene in the mosquito. We show that AHR3 is expressed in both vitellogenic tissues of the female mosquito, the fat body and the ovary. The expression of AHR3 correlates with the ecdysteroid titer, reaching a peak at 24 h after a blood meal. Moreover, in vitro fat body culture experiments demonstrate that the kinetics and dose response of AHR3 to 20-hydroxyecdysone (20E), an active ecdysteroid in the mosquito, is similar to those of the late vitellogenic genes rather than the early E75 gene. However, as shown for other early and early-late genes, the 20E activation of AHR3 is not inhibited by the presence of cycloheximide, a protein synthesis inhibitor. Taken together, these findings strongly suggest AHR3 involvement in regulating the vitellogenic response to ecdysone in the adult mosquito.     

Correlation of human epidermal growth factor receptor 2 expression with clinicopathological characteristics and prognosis in gastric cancer

AIM:To investigate human epidermal growth factor receptor 2(HER2) gene amplification and protein expression in Chinese patients with resectable gastric cancer and the association with clinicopathological characteristics and survival.METHODS:One hundred and ninety-seven gastric cancer patients who underwent curative surgery procedures were enrolled into this study.HER2 gene amplification and protein expression were examined using fluorescence in-situ hybridization(FISH) and immunohistochemistry(IHC) analysis on formalin-fixed paraffinembedded gastric cancer samples from all patients.For scoring,Hofmann’s HER2 gastric cancer scoring system was adopted.All cases showing IHC3+ or FISH positiv-ity were defined as HER2 positive.Patient clinicopathological data and survival information were collected.Finally,χ 2 statistical analysis was performed to analyze the HER2 positivity rate amongst the subgroups with different clinicopathological characteristics including;gender,age,tumor location,Lauren classification,differentiation,TNM staging,depth of invasion,lymph node metastases and distant metastasis.The probability of survival for different subgroups with different clinicopathological characteristics was calculated using the Kaplan-Meier method and survival curves plotted using log rank inspection.RESULTS:According to Hofmann’s HER2 gastric cancer scoring criteria,31 cases(15.74%) were identified as HER2 gene amplified and 19 cases(9.64%) were scored as strongly positive for HER2 membrane staining(3+),25 cases(12.69%) were moderately positive(2+) and 153 cases(77.66%) were HER2 negative(0/1+).The concordance rate between IHC and FISH analyses was 88.83%(175/197).Thirty-six cases were defined as positive for HER2 gene amplification and/or protein expression,with 24 of these cases being eligible for Herceptin treatment according to United States recommendations,and 29 of these cases eligible according to EU recommendations.Highly consistent results were detected between IHC3+,IHC0/1 and FISH(73.68% and 95.42%),but l     

广西壮族自治区耐多药结核分枝杆菌耐药情况与基因型特征分析

目的 了解广西壮族自治区(简称“广西”)耐多药结核分枝杆菌(MDR-MTB)的耐药情况、基因型构成及基因型与耐药的相关性,为耐多药结核病的防控提供理论依据。方法 采用连续监测的方法,选取位于广西境内东、西、南、北、中的贵港、百色、崇左、桂林和防城港5个市为监测点,采用随机数字表法抽取5个市中的21个县(市、区),纳入于2016—2017年在当地结核病防治(简称“结防”)机构登记治疗且培养阳性的MTB菌株共1514株,使用WHO推荐的比例法对异烟肼(INH)、利福平(RFP)、乙胺丁醇(EMB)、链霉素(Sm)、氧氟沙星(Ofx)和卡那霉素(Km)进行耐药性检测,最终有51株确定为MDR-MTB菌株。运用熔解曲线间隔区寡核苷酸分型法(McSpoligotyping)对MDR-MTB菌株进行基因分型,将分型结果与SpolDB4.0数据库进行比对。结果 51株MDR-MTB菌株对EMB、Sm、Ofx和Km的耐药率分别为41.18%(21/51)、31.37%(16/51)、9.80%(5/51)和1.96%(1/51)。北京基因型菌株占56.86%(29/51),非北京基因型菌株占43.14%(22/51)。对EMB耐药的菌株中,北京基因型14株,占66.67%(14/21),非北京基因型7株,占33.33%(7/21),差异无统计学意义(χ ²=1.399,P=0.237);对Sm耐药的菌株中,北京基因型11株,占68.75%(11/16),非北京基因型5株,占31.25%(5/16),差异无统计学意义(χ ²=1.343,P=0.246);对Ofx耐药的菌株中,北京基因型9株,占60.00%(9/15),非北京基因型6株,占40.00%(6/15),差异无统计学意义(χ ²=0.085,P=0.770);对Km耐药的菌株中,北京基因型1株,占100.00%(1/1),非北京基因型0株,占0.00%(0/1),差异无统计学意义(P=1.000)。结论 应重视广西MDR-MTB菌株对EMB、Sm、Ofx和Km的耐药情况;MDR-MTB菌株主要为北京基因型;北京和非北京基因型对EMB、Sm、Ofx和Km的耐药率未见差异。