BACKGROUND & AIMS: -2

背景与目标： -2

BACKGROUND & AIMS: :Pseudomonas aeruginosa is a gram-negative opportunistic pathogen that is cytotoxic towards a variety of eukaryotic cells. To investigate the effect of this bacterium on monocyte, we infected human U937 cells with the P. aeruginosa strain in vitro. To explore the expression of Bcl-2 and Bax as well as caspase-3/9 activation in the apoptosis of human U937 cells induced by P. aeruginosa, Hoechst 33258 staining and Giemsa staining as well as Flow cytometry analysis were used to determine the rate of apoptosis, and the expressions of Bcl-2 and Bax were assayed by RT-PCR and Western blotting respectively. Bax protein conformation change was assayed by immunoprecipitation. Cytochrome c release was measured by Western blotting. Moreover, exposure of U937 cells to P. aeruginosa measured caspase-3/9 activity. It was found that the apoptosis of human U937 cells could be induced by Pseudomonas aeruginosa in a dose- and time-dependent manner. Also, there were a tendency of alterations with an increased expression level of Bax and a reduced expression level of Bcl-2, increased levels of cytochrome c release, and also with an increased activation of caspase-3/9 and Bax protein conformation change. For the evaluation of the role of caspases, caspase-3/9 inhibitors Z-DEVD-FMK and Z-LEHD-FMK respectively were used. The results were further confirmed by the observation that the caspase inhibitors Z-DEVD-FMK and Z-LEHD-FMK blocked P. aeruginosa-induced U937 apoptosis. It is concluded that P. aeruginosa can induce apoptosis with an up-regulated expression of Bax and a down-regulated expression of Bcl-2, which resulted in increased levels of cytochrome c release and increased caspase-3 and -9 in human U937 cells.

背景与目标： 铜绿假单胞菌是革兰氏阴性机会病原体，对多种真核细胞具有细胞毒性。为了研究这种细菌对单核细胞的作用，我们在体外用铜绿假单胞菌菌株感染了人U937细胞。为了探索铜绿假单胞菌诱导的人U937细胞凋亡中Bcl-2和Bax的表达以及caspase-3 / 9的激活，使用Hoechst 33258染色和Giemsa染色以及流式细胞仪分析来确定其发生率。分别用RT-PCR和Western blotting检测Bcl-2和Bax的表达。通过免疫沉淀测定Bax蛋白的构象变化。通过蛋白质印迹法测定细胞色素c的释放。此外，U937细胞暴露于铜绿假单胞菌可测量caspase-3 / 9活性。发现铜绿假单胞菌可以剂量和时间依赖性诱导人U937细胞的凋亡。同样，Bax表达水平升高，Bcl-2表达水平降低，细胞色素c释放水平升高，caspase-3 / 9激活激活和Bax蛋白构象变化也存在改变的趋势。为了评估胱天蛋白酶的作用，分别使用了caspase-3 / 9抑制剂Z-DEVD-FMK和Z-LEHD-FMK。通过观察到胱天蛋白酶抑制剂Z-DEVD-FMK和Z-LEHD-FMK阻断了铜绿假单胞菌诱导的U937细胞凋亡，进一步证实了该结果。结论：铜绿假单胞菌可以诱导凋亡，其Bax表达上调而Bcl-2表达下调，从而导致人U937细胞中细胞色素c的释放水平升高以及caspase-3和-9升高。 。

BACKGROUND & AIMS: :Reproductive follow-up of carriers of familial reciprocal balanced translocations involving chromosome 9 and comparison with predicted outcome: Chromosome 9 is commonly implicated in reciprocal translocations (rcp). Twenty-seven families segregating rcp involving chromosome 9 were selected with the aim of comparing the theoretical risk of Mental Retardation with Congenital Anomalies (MCA/MR) calculated according to Human Cytogenetics Forum with the observed reproductive follow-up. The 27 families include 157 subjects. The reproductive follow-up showed that the majority of mothers underwent full-term pregnancies (88/130), and that there were 37 spontaneous and five voluntary abortions. Eighty-one subjects were karyotyped: 18 had a normal karyotype, 50 carried an rcp, ten had an unbalanced rcp-related karyotype and three an abnormal rcp-unrelated karyotype. Of the 88 live-born individuals, seven had an abnormal rcp-related karyotype with partial chromosome 9 trisomy (four cases) or partial 9p monosomy (three cases), and 48 were rcp carriers, two of whom also presented additional anomalies. The evaluation of reproductive outcomes in the 27 families studied revealed good concordance between the Human Cytogenetics Forum predictions and the observed follow-up in relation to the most probable mode of unbalance at birth, and the higher risk of MCA/MR in rcp carriers with unbalanced live-borns in comparison with those generating healthy progeny

背景与目标： ：涉及9号染色体的家族相互平衡易位的携带者的生殖随访并与预期结果进行比较：9号染色体通常与相互易位（rcp）有关。选择了27个涉及9号染色体的rcp分离家庭，目的是将根据人类细胞遗传学论坛计算的先天性精神发育迟滞的理论风险与观察到的生殖随访进行比较。 27个家庭包括157个科目。生殖随访表明，大多数母亲都进行了足月妊娠（88/130），并且有37例自然流产和5例自愿流产。八十一名受试者进行了核型分析：18名受试者具有正常的核型，50名受试者具有rcp，十名受试者具有不平衡的rcp相关性核型，三名受试者具有异常的rcp无关性核型。在88个活产个体中，有7个具有与rcp相关的核型异常，其中9号染色体三体性（4例）或9p单体性染色体3个（3例），并且rcp携带者48个，其中两个还存在其他异常。对所研究的27个家庭的生殖结果进行的评估表明，人类细胞遗传学论坛的预测与观察到的关于出生时最可能的失衡方式以及在失衡的rcp携带者中MCA / MR的风险较高之间的随访之间具有良好的一致性与产生健康后代的活产婴儿相比

BACKGROUND & AIMS: :Toll like receptors (TLRs) are essential molecules implicated in both innate and adaptive immune response. Polymorphisms in TLR gene have been associated with various infectious diseases and autoimmune disorders. Role of TLR9 has been elegantly demonstrated in both human systemic lupus erythematosus (SLE) and mice model of lupus. In the present study we investigated association of TLR-9 promoter polymorphisms (T-1237C and T-1486C) with susceptibility/resistance to SLE in an Eastern Indian state which is endemic to parasitic diseases. 210 Female SLE patients who fulfilled the American College of Rheumatology criteria were enrolled along with matched healthy controls from Odisha, India. TLR-9 polymorphisms (T-1237C and T-1486C) were typed by polymerase chain reaction followed by restriction fragment length polymorphism. For meta-analysis, relevant literatures were searched from PubMed database and comprehensive meta-analysis V2 software was employed for analysis. Allele and genotype frequency of TLR-9 promoter polymorphisms (T-1237C and T-1486C) were comparable among SLE patients and controls. Further, meta-analysis of earlier reports and present study did not reveal a significant association of TLR-9 (T-1237C and T-1486C) polymorphisms with SLE. Data from the present study suggest that TLR-9 promoter polymorphisms are not associated with susceptibility to SLE in an area endemic to parasitic diseases.

背景与目标： ：Toll样受体（TLRs）是与先天和适应性免疫反应有关的必需分子。 TLR基因的多态性与多种传染病和自身免疫性疾病有关。 TLR9的作用已在人系统性红斑狼疮（SLE）和狼疮小鼠模型中得到了很好的证明。在本研究中，我们调查了在印度洋东部寄生虫病流行州，TLR-9启动子多态性（T-1237C和T-1486C）与对SLE的敏感性/耐药性之间的关系。纳入了210名符合美国风湿病学会标准的女性SLE患者以及来自印度奥里萨邦的相匹配的健康对照。 TLR-9多态性（T-1237C和T-1486C）通过聚合酶链式反应，然后进行限制性片段长度多态性进行分型。对于荟萃分析，从PubMed数据库中搜索相关文献，并使用综合的荟萃分析V2软件进行分析。在SLE患者和对照组中，TLR-9启动子多态性（T-1237C和T-1486C）的等位基因和基因型频率相当。此外，对早期报道和本研究的荟萃分析未发现TLR-9（T-1237C和T-1486C）多态性与SLE有显着相关性。来自本研究的数据表明，在寄生虫病流行地区，TLR-9启动子多态性与SLE易感性无关。

BACKGROUND & AIMS: BACKGROUND:Meta-analyses enable summarization and interpretation of data across clinical trials. When applied to safety data they allow for detection of rare events. Recently, a 13-valent pneumococcal conjugate vaccine (PCV13) was approved in multiple countries worldwide for routine immunization of infants and young children. This meta-analysis was conducted to identify potentially clinically important rare safety events associated with PCV13. OBJECTIVE:To summarize the safety of PCV13 compared with 7-valent pneumococcal conjugate vaccine (PCV7) administered to infants and toddlers. METHODS:A meta-analysis was performed of integrated safety data from 13 infant studies (PCV13 n=4729 and PCV7 n=2760) conducted in 9 North American, European, and Asian countries. Local reactions at the vaccine injection site and systemic events were collected for 4-7 days after each dose into electronic diaries. Adverse events (AEs) were collected after each vaccination. RESULTS:Overall, rates of local reactions after any dose of the infant series were similar between PCV13 and PCV7 groups: tenderness (46.7% vs 44.8%, respectively); swelling (28.5% vs 26.9%); and redness (36.4% vs 33.9%). After the toddler dose, tenderness was significantly higher among PCV7 subjects than PCV13 subjects (54.4% vs 48.8%; P=0.005). Frequencies of fever (≥38°C) were similar in both groups and mostly mild (≤39°C); incidence of moderate fever (>39°C to ≤40°C) with PCV13 was ≤2.8% after any infant dose and 5.0% after the toddler dose, compared with ≤2.6% and 7.3%, respectively, with PCV7. Fever >40°C was uncommon in both groups. Frequencies of decreased appetite, irritability, and sleep disturbances were similar in both groups. AEs were the types of conditions and symptoms expected in infants and children, and clinically significant differences between vaccine groups were not observed. CONCLUSION:PCV13 has a favorable safety profile similar to that of PCV7, a vaccine for which there is >10 years clinical experience.

背景与目标： 背景：元分析可以汇总和解释整个临床试验中的数据。当应用于安全数据时，它们允许检测罕见事件。最近，一种13价肺炎球菌结合疫苗（PCV13）已在全球多个国家/地区批准用于婴儿和幼儿的常规免疫。进行了这项荟萃分析，以鉴定与PCV13相关的潜在的临床重要罕见安全事件。
目的：总结与婴幼儿使用的7价肺炎球菌结合疫苗（PCV7）相比，PCV13的安全性。
方法：对9个北美，欧洲和亚洲国家进行的13项婴儿研究（PCV13 n = 4729和PCV7 n = 2760）的综合安全性数据进行了荟萃分析。每次注射疫苗后4-7天，将疫苗注射部位的局部反应和全身性事件收集到电子日记中。每次疫苗接种后收集不良事件（AE）。
结果：总的来说，在PCV13和PCV7组中，任何剂量的婴儿系列后的局部反应率相似：压痛（分别为46.7％和44.8％）；肿胀（28.5％对26.9％）;和发红（36.4％比33.9％）。幼儿服药后，PCV7受试者的压痛明显高于PCV13受试者（54.4％比48.8％； P = 0.005）。两组的发烧频率（≥38°C）相似，且多数为轻度（≤39°C）。在任何婴儿剂量下，PCV13发生中度发烧（> 39°C到≤40°C）的发生率≤2.8％，在幼儿剂量后为5.0％，而PCV7分别为≤2.6％和7.3％。两组中发烧> 40°C的情况很少见。两组食欲减退，烦躁不安和睡眠障碍的频率相似。不良事件是婴儿和儿童中预期的疾病和症状类型，未观察到疫苗组之间的临床显着差异。
结论：PCV13具有与PCV7类似的良好安全性，后者具有10年以上的临床经验。

BACKGROUND & AIMS: :Cellular-FLICE inhibitory protein (c-FLIP) is an inhibitor of apoptosis downstream of the death receptors Fas, DR4, and DR5, and is expressed as long (c-FLIP(L)) and short (c-FLIP(S)) splice forms. We found that the knockdown of c-FLIP using small interfering RNA (siRNA) triggered ligand-independent caspase-8- and -9-dependent spontaneous apoptosis and decreased the proliferation of MCF-7 breast cancer cells. Further analysis revealed that an apoptotic inhibitory complex (AIC) comprised of DR5, FADD, caspase-8, and c-FLIP(L) exists in MCF-7 cells, and the absence of c-FLIP(L) from this complex induces DR5- and FADD-mediated caspase-8 activation in the death inducing signaling complex (DISC). c-FLIP(S) was not detected in the AIC, and using splice form-specific siRNAs we showed that c-FLIP(L) but not c-FLIP(S) is required to prevent spontaneous death signaling in MCF-7 cells. These results clearly show that c-FLIP(L) prevents ligand-independent death signaling and provides direct support for studying c-FLIP as a relevant therapeutic target for breast cancers.

背景与目标： ：细胞FLICE抑制蛋白（c-FLIP）是死亡受体Fas，DR4和DR5下游的凋亡抑制剂，表达为长（c-FLIP（L））和短（c-FLIP（S） ）拼接形式。我们发现使用小干扰RNA（siRNA）敲低c-FLIP会触发不依赖配体的caspase-8和-9依赖性自发凋亡，并降低MCF-7乳腺癌细胞的增殖。进一步的分析表明，MCF-7细胞中存在由DR5，FADD，caspase-8和c-FLIP（L）组成的凋亡抑制复合物（AIC），并且该复合物中不存在c-FLIP（L）会诱导DR5。 -和FADD介导的caspase-8激活在死亡诱导信号复合物（DISC）中。在AIC中未检测到c-FLIP（S），使用剪接形式特异性siRNA，我们发现c-FLIP（L）而非c-FLIP（S）是防止MCF-7细胞自发死亡信号所必需的。这些结果清楚地表明，c-FLIP（L）可以防止配体依赖性死亡信号传导，并为研究c-FLIP作为乳腺癌的相关治疗靶标提供了直接支持。

BACKGROUND & AIMS: :The mammalian matrix metalloproteinase-9 (MMP-9), which might play a role in ovulation, uterus remodeling, embryo development, and implantation in mammals, is one of the potential functional candidate genes for porcine reproductive traits. In this study, the entire genomic sequence of porcine MMP-9 (pMMP-9) gene was established; it contains 13 exons and 12 introns. Real-time PCR analysis revealed that pMMP-9 is highly expressed in the Minzhu uterus before puberty and decreases significantly after sexual maturity (p < 0.05). Two single-nucleotide polymorphisms (A3011G and T5079C) that can be detected by PCR restriction fragment length polymorphism (PCR-RFLP) were discovered and tested for statistical associations with litter size traits in a crossbred population (Line DIV) derived from Landrace, Large White, Chinese Tongcheng and/or Chinese Meishan pigs. For A3011G, the GG genotype was associated with a significantly higher (p < 0.05) number of live births than those recorded for AA sows and the additive effect was significant (p < 0.05). The T5079C marker is not associated with litter size in this population. Further studies are needed to confirm the results of this study.

背景与目标： ：哺乳动物基质金属蛋白酶9（MMP-9），可能在排卵，子宫重塑，胚胎发育和哺乳动物的植入中起作用，是猪繁殖性状的潜在功能候选基因之一。在这项研究中，建立了猪MMP-9（pMMP-9）基因的完整基因组序列。它包含13个外显子和12个内含子。实时PCR分析显示，pMMP-9在青春期前的zhu竹子宫中高表达，在性成熟后明显降低（p <0.05）。发现了两个可以通过PCR限制性片段长度多态性（PCR-RFLP）检测到的单核苷酸多态性（A3011G和T5079C）并测试了来自长白大白杂交种（Line DIV）与杂种大小性状的统计关联，中国桐城猪和/或中国眉山猪。对于A3011G，GG基因型的活产数显着高于（AA）母猪记录的活产数（p <0.05），累加效应显着（p <0.05）。 T5079C标记与该种群中的产仔数无关。需要进一步的研究以确认这项研究的结果。

BACKGROUND & AIMS: :The structural and functional analyses of heterochromatin are essential to understanding how heterochromatic genes are regulated and how centromeric chromatin is formed. Because the repetitive nature of heterochromatin hampers its genome analysis, new approaches need to be developed. Here, we describe how, in double mutants for Su(var)3-9 and SuUR genes encoding two structural proteins of heterochromatin, new banded heterochromatic segments appear in all polytene chromosomes due to the strong suppression of under-replication in pericentric regions. FISH on salivary gland polytene chromosomes from these double mutant larvae allows high resolution of heterochromatin mapping. In addition, immunostaining experiments with a set of antibodies against euchromatic and heterochromatic proteins reveal their unusual combinations in the newly appeared segments: binding patterns for HP1 and HP2 are coincident, but both are distinct from H3diMetK9 and H4triMetK20. In several regions, partial overlapping staining is observed for the proteins characteristic of active chromatin RNA Pol II, H3triMetK4, Z4, and JIL1, the boundary protein BEAF, and the heterochromatin-enriched proteins HP1, HP2, and SU(VAR)3-7. The exact cytological position of the centromere of chromosome 3 was visualized on salivary gland polytene chromosomes by using the centromeric dodeca satellite and the centromeric protein CID. This region is enriched in H3diMetK9 and H4triMetK20 but is devoid of other proteins analyzed.

背景与目标： ：异染色质的结构和功能分析对于了解异色基因如何调控以及着丝粒染色质如何形成至关重要。由于异染色质的重复性质妨碍了其基因组分析，因此需要开发新的方法。在这里，我们描述了如何在Su（var）3-9和SuUR基因的双突变体中编码异染色质的两个结构蛋白，新条带化的异色片段由于强烈抑制外周中心区域的复制不足而在所有多烯染色体中出现。这些双突变幼虫在唾液腺多态染色体上的FISH可以实现高分辨率的异染色质定位。此外，使用针对常染色体和异源蛋白质的抗体的免疫染色实验在新出现的片段中揭示了它们的不寻常组合：HP1和HP2的结合模式是重合的，但两者均不同于H3diMetK9和H4triMetK20。在几个区域中，观察到活性染色质RNA Pol II，H3triMetK4，Z4和JIL1，边界蛋白BEAF和富含异染色质的蛋白HP1，HP2和SU（VAR）3-7的蛋白具有部分重叠染色。 。通过使用着丝粒十二指肠卫星和着丝粒蛋白CID，在唾液腺多烯染色体上可以看到3号染色体着丝粒的确切细胞学位置。该区域富含H3diMetK9和H4triMetK20，但没有其他被分析的蛋白质。

BACKGROUND & AIMS: Background:Childhood non-vaccination can have different short-and long-term negative outcomes on their health. In Ethiopia, in addition to low coverage of full vaccination, street children were among the neglected part of the community who were missed during planning and reporting vaccination coverage. Moreover, there is no related research conducted on this title specifically. Objective:The objective of the study was to assess the vaccination status and its associated factors among street children 9-24 months old in Sidama zone. Methods:Community-based cross-sectional study design was conducted in four selected towns of Sidama region, southern Ethiopia. The convenience sampling method was applied to involve mothers of street children younger than two years during the study period. Data entry was done with EpiData version 3.1 and exported to SPSS22 for analysis. Bivariate and multivariable logistic regression analysis were performed to identify factors associated with immunization status of street children. Results:A significant number (26 [24.3%]) of the street children younger than two years were not vaccinated. Those mothers who are ≤20 years old (P = 0.014, AOR = 0.216, 95% CI: 0.064-0.732) and who gave birth at home (P = 0.029, AOR = 0.292, 95% CI: 0.097-0.879) had less odds of vaccinating their child than those older than 20 and who gave birth at health facility respectively. Conclusion:A significant number of the street children in this study are not fully vaccinated. Mothers aged <20 years and home births were significantly associated with non-vaccination status.

背景与目标： 背景：儿童未接种疫苗可能会对他们的健康产生不同的短期和长期负面影响。在埃塞俄比亚，除了全面接种疫苗的覆盖率低外，流浪儿童是社区中被忽略的一部分，在计划和报告接种疫苗覆盖率时被遗漏了。而且，还没有对此标题进行专门的相关研究。
目的：本研究的目的是评估西达玛州9-24个月大的街头儿童的疫苗接种状况及其相关因素。
方法：在埃塞俄比亚南部锡达玛地区的四个选定城镇中进行了基于社区的横断面研究设计。在研究期间，便利抽样方法被用于涉及不到两岁的流浪儿童的母亲。数据输入使用EpiData 3.1版完成，并导出到SPSS22进行分析。进行了双变量和多变量logistic回归分析，以确定与流浪儿童免疫状况相关的因素。
结果：相当多（26 [24.3％]）岁以下的街头儿童未接种疫苗。 ≤20岁（P = 0.014，AOR = 0.216，95％CI：0.064-0.732）且在家分娩（P = 0.029，AOR = 0.292，95％CI：0.097-0.879）的母亲较少给他们的孩子接种疫苗的几率分别高于20岁和在卫生机构分娩的孩子。
结论：本研究中有相当数量的流浪儿童没有完全接种疫苗。年龄小于20岁的母亲和家庭生育与非疫苗接种状态显着相关。

BACKGROUND & AIMS: BACKGROUND:The balance between proteinases and antiproteinases plays an important role in tissue destruction and remodelling. In chronic obstructive pulmonary disease (COPD) and emphysema, an imbalance between matrix metalloproteinases (MMPs) and inhibitors of tissue metalloproteinase (TIMPs) has been reported. Alveolar macrophages are considered to be the main source of MMPs. We therefore have analyzed the effects of free and liposomal all trans-retinoic acid (ATRA) on the expression of MMP-9 and TIMP-1 in bronchoalveolar lavage (BAL) cells from patients with COPD and patients with other lung diseases. MATERIAL AND METHODS:BAL cells were incubated 1-3 day with either liposomal or free ATRA. Supernatants were tested for MMP-9 and TIMP-1 protein in specific ELISA systems; mRNA analysis was performed by semi-quantitative RT-PCR and by quantitative LightCycler PCR. RESULTS:We demonstrate that either liposomal or free ATRA selectively down-regulates MMP-9 and up-regulates TIMP-1. At the protein level, MMP-9 is decreased 3-fold and TIMP-1 is increased 3.5-fold compared to the base line with empty liposomes or untreated cells. The ratio of MMP-9 and its inhibitor TIMP-1, which may be crucial to the overall proteolytic potential decreased by factor 8. That this countercurrent effect of ATRA is not due to an altered protein stability but to transcriptional regulation could be demonstrated by RT-PCR. Quantitative LightCycler analysis revealed a 2.5-fold decrease of MMP-9 mRNA and a 4.5 fold increase of TIMP- 1 mRNA. CONCLUSIONS:These data suggest that ATRA treatment via its impact on the proteinase/antiproteinase ratio may become a new therapeutic strategy for patients with inflammatory destructive lung diseases.

背景与目标： 背景：蛋白酶和抗蛋白酶之间的平衡在组织破坏和重塑中起着重要作用。在慢性阻塞性肺疾病（COPD）和肺气肿中，基质金属蛋白酶（MMPs）和组织金属蛋白酶抑制剂（TIMPs）之间存在失衡的报道。肺泡巨噬细胞被认为是MMP的主要来源。因此，我们分析了游离和脂质体全反式维甲酸对COPD患者和其他肺部疾病患者支气管肺泡灌洗（BAL）细胞中MMP-9和TIMP-1表达的影响。
材料与方法：将BAL细胞与脂质体或游离ATRA孵育1-3天。在特定的ELISA系统中检测上清液中的MMP-9和TIMP-1蛋白。通过半定量RT-PCR和定量LightCycler PCR进行mRNA分析。
结果：我们证明脂质体或游离ATRA选择性下调MMP-9和上调TIMP-1。与空脂质体或未经处理的细胞相比，在蛋白质水平上，MMP-9降低了3倍，TIMP-1升高了3.5倍。 MMP-9及其抑制剂TIMP-1的比例可能对总蛋白水解潜力至关重要，降低了8倍。ATRA的这种逆流作用不是由于蛋白质稳定性的改变，而是转录调控，可以通过RT证实。 -PCR。定量LightCycler分析显示MMP-9 mRNA下降了2.5倍，TIMP-1 mRNA上升了4.5倍。
结论：这些数据表明，通过对蛋白酶/抗蛋白酶比率的影响，ATRA治疗可能成为炎性破坏性肺疾病患者的一种新的治疗策略。

BACKGROUND & AIMS: :The multidrug resistant cell line DC-3F/ADII was obtained by stepwise selection for growth in actinomycin D (ActD). Compared with parental cells, it displays high resistance to ActD and vincristine and low resistance to colchicine and daunorubicin. These cells overexpress a form of P-glycoprotein (Pgp1) containing a double mutation, I837L and N839I, in transmembrane domain (TM) 9; when transfected into DC-3F, this mutation confers the DC-3F/ADII phenotype. We have shown previously that another cell line, DC-3F/ADX, also displays this phenotype and overexpresses a mutant form of Pgp1 containing a double mutation in TM6 (G338A, A339P). Hence, mutations in TM9 and TM6 are independently capable of conferring the same cross-resistance phenotype. The TM6 mutations inhibit the ability of cyclosporin A to reverse cross-resistance and to block labeling of the protein by [125I]iodoarylazidoprazosin (IAAP), whereas the TM9 mutations do not show similar effects. A chimeric protein containing both pairs of mutations confers twice the level of resistance to ActD than expected from the sum of the individual mutations, but it cannot be labeled to detectable levels with [125I]IAAP. Thus, TM9 represents a novel site that cooperates with TM6 to mediate drug resistance and [125I]IAAP labeling.

背景与目标： ：通过逐步选择放线菌素D（ActD）中的生长获得多药耐药细胞系DC-3F / ADII。与亲代细胞相比，它对ActD和长春新碱具有较高的抵抗力，而对秋水仙碱和柔红霉素的抵抗力则较低。这些细胞过表达一种P-糖蛋白（Pgp1），该蛋白在跨膜结构域（TM）9中包含一个双突变，即I837L和N839I。当转染至DC-3F时，此突变赋予DC-3F / ADII表型。先前我们已经证明，另一种细胞系DC-3F / ADX也显示此表型，并过表达Pgp1的突变形式，该突变形式包含TM6中的双重突变（G338A，A339P）。因此，TM9和TM6中的突变能够独立赋予相同的交叉抗性表型。 TM6突变抑制环孢菌素A反向交叉耐药性并阻止[125I]碘代芳基叠氮吡唑嗪（IAAP）标记蛋白质的能力，而TM9突变没有显示出相似的作用。包含两对突变的嵌合蛋白对ActD的抗性水平要比单个突变的总和预期的抗性水平高一倍，但无法用[125I] IAAP标记为可检测水平。因此，TM9代表一个与TM6协同介导耐药性和[125I] IAAP标记的新位点。

BACKGROUND & AIMS: BACKGROUND:IL-9 is an important cytokine in allergic diseases such as asthma, atopic dermatitis, etc. T helper (Th) cells seem to be the main source of IL-9. Cellular and molecular mechanisms of IL-9 production by human Th cells have been poorly understood. METHODS:Dermatophagoides farinae(Der f)-specific Th clones were established from peripheral blood lymphocytes of atopic asthmatics, and cytokine synthesis in response to various stimuli was determined by specific ELISAs. RESULTS:IL-9 was produced by 14 of 27 human Th clones upon T cell receptor (TCR) stimulation, immobilized anti-CD3 antibody (Ab). IL-9 production was significantly enhanced by the addition of anti-CD28 Ab into the culture, indicating the role of costimulatory signal on IL-9 synthesis. Pharmacologically, IL-9 production was induced by ionomycin (IOM) alone, and enhanced by phorbol 12-myristate 13-acetate (PMA). rIL-2 induced IL-9 production by 8 out of 19 Th clones. IL-9 production by Th clones stimulated with immobilized anti-CD3 Ab was significantly suppressed by the addition of anti-IL-2 neutralizing Ab into the culture. CONCLUSION:Approximately half of the Der f-specific Th clones derived from atopic asthmatics produced IL-9 upon TCR stimulation. Ca(2+) signal, CD28 signal, and IL-2 receptor signal seem to play important roles in IL-9 production by human Th cells. Moreover, synthesis of IL-9, a Th2 cytokine, is dependent on IL-2, a Th1 cytokine, which is produced by Th cells themselves.

背景与目标： 背景：IL-9在过敏性疾病（例如哮喘，特应性皮炎等）中是重要的细胞因子。T辅助（Th）细胞似乎是IL-9的主要来源。人们对人Th细胞产生IL-9的细胞和分子机制了解甚少。
方法：从特应性哮喘患者外周血淋巴细胞中建立特异的Dermatophagoides farinae（Der f）Th克隆，并通过特异性ELISA法测定响应各种刺激的细胞因子合成。
结果：IL-9由27个人类Th克隆中的14个在T细胞受体（TCR）刺激下产生，并固定了抗CD3抗体（Ab）。向培养物中添加抗CD28 Ab可显着增强IL-9的产生，表明共刺激信号对IL-9合成的作用。在药理上，IL-9的产生仅由离子霉素（IOM）诱导，并由佛波12-肉豆蔻酸酯13-乙酸酯（PMA）增强。 rIL-2诱导19个克隆中的8个产生IL-9。通过向培养物中添加抗IL-2中和抗体，显着抑制了由固定化抗CD3 Ab刺激的Th克隆产生的IL-9。
结论：来自特应性哮喘的Der f-特异性Th克隆约有一半在TCR刺激下产生IL-9。 Ca（2）信号，CD28信号和IL-2受体信号似乎在人类Th细胞产生IL-9中起重要作用。此外，IL-9（Th2细胞因子）的合成依赖于Th-2细胞本身产生的IL-2（Th1细胞因子）。

BACKGROUND & AIMS: :Early detection of polyps or colorectal carcinoma can reduce colorectal carcinoma-associated deaths. Previous studies have demonstrated raised serum levels of matrix metalloproteinase 9 (sMMP-9) in a range of cancers. The aim of this study was to investigate the role of sMMP-9 levels in identifying colorectal neoplasia. Consenting patients donated a blood sample and were assessed by proforma-led history and physical examination. Samples were analysed for sMMP-9 concentration (enzyme-linked immuno-sorbant assay) and compared to final diagnoses. Logistic regression modelling determined independent factors associated with neoplasia. A total of 365 patients were recruited of whom 300 were analysed, including 46 normal controls. A total of 27 significant adenomas and 63 malignancies were identified. The median sMMP-9 concentration was 443 ng ml(-1) (IQR: 219-782; mean: 546). Patients with neoplasia had significantly elevated sMMP-9 levels (P<0.001). Logistic regression modelling identified elevated log(sMMP-9) as the most significant predictor of neoplasia (chi(2)=38.33, P<0.001). Other significant factors were age, sex, smoking history, abdominal pain and weight loss. The model accurately predicted neoplasia in 77.3% of cases. Sensitivity and specificity were 77.9 and 77.1%. sMMP-9 estimation can accurately stratify patient to low- or high-risk cohorts. Serum sampling is a potential means of avoiding unnecessary colonoscopy and reducing patient anxiety, iatrogenic morbidity and mortality, and cost.

背景与目标： ：及早发现息肉或大肠癌可以减少与大肠癌相关的死亡。先前的研究表明，在多种癌症中血清基质金属蛋白酶9（sMMP-9）的血清水平升高。这项研究的目的是调查sMMP-9水平在鉴定结直肠肿瘤中的作用。同意的患者捐赠了血液样本，并通过形式记录的病史和体格检查进行了评估。分析样品的sMMP-9浓度（酶联免疫吸附测定），并与最终诊断结果进行比较。 Logistic回归模型确定了与瘤形成相关的独立因素。总共招募了365位患者，其中300位被分析，包括46位正常对照。总共鉴定出27个重要的腺瘤和63个恶性肿瘤。 sMMP-9的中位数浓度为443 ng ml（-1）（IQR：219-782;平均值：546）。瘤形成患者的sMMP-9水平显着升高（P <0.001）。 Logistic回归模型确定升高的log（sMMP-9）是瘤形成的最重要预测因子（chi（2）= 38.33，P <0.001）。其他重要因素是年龄，性别，吸烟史，腹痛和体重减轻。该模型在77.3％的病例中可以准确预测肿瘤。敏感性和特异性分别为77.9％和77.1％。 sMMP-9评估可以将患者分为低危或高危人群。血清采样是避免不必要的结肠镜检查并减少患者焦虑，医源性发病率和死亡率以及成本的潜在手段。

BACKGROUND & AIMS: :Congo red (CR) is a commonly used histological amyloid dye and a weak amyloid inhibitor. There is currently no experimentally available structure of CR bound to an amyloid fibril and the binding modes, and the mechanisms governing its inhibitory and optical properties are poorly understood. In this work, we present the first, to our knowledge, atomistically detailed picture of CR binding to protofibrils of the Alzheimer Aβ(9-40) peptide. We identify three major binding modes, with the primary mode residing in the grooves formed by the β-sheets, and observe a restriction of the torsional rotation of the CR molecule upon binding. Our simulations reveal a novel, to our knowledge, electrostatic steering mechanism that plays an important role in the initial recognition and binding of CR to the positively charged surface residues of the fibril. Our simulations provide new, to our knowledge, insights into the striking spectrophotometric and inhibitory properties of CR. In particular, we show that birefringence upon CR binding is due to the anisotropic orientation of the CR dipoles resulting from the spatial ordering of these molecules in the grooves along the fibril axis. The fluorescent enhancement of the bound CR, in turn, is associated with the torsional restriction of this molecule upon binding.

背景与目标： 刚果红（CR）是常用的组织学淀粉样蛋白染料和弱淀粉样蛋白抑制剂。目前尚没有实验上可得到的与淀粉样蛋白原纤维结合的CR结构和结合方式，而对其抑制和光学性质的控制机理知之甚少。在这项工作中，据我们所知，我们首次呈现了与阿尔茨海默病Aβ（9-40）肽原纤维结合的CR的原子学详细图片。我们确定了三种主要的结合模式，主要模式驻留在由β-折叠形成的凹槽中，并观察到结合后CR分子扭转旋转的限制。我们的模拟揭示了一种据我们所知的新型静电操纵机制，该机制在CR的初次识别和与原纤维表面带正电荷的残基的结合中起着重要作用。据我们所知，模拟为CR的惊人的分光光度法和抑制性质提供了新的见解。特别地，我们表明，CR结合的双折射归因于CR偶极子的各向异性取向，这是由于这些分子在沿原纤维轴的凹槽中的空间顺序所致。结合的CR的荧光增强又与该分子在结合时的扭转限制有关。

BACKGROUND & AIMS: :Wet deposition of nitrogen (N) occurs in oxidized (nitrate) and reduced (ammonium) forms. Whether one form drives vegetation change more than the other is widely debated, as field evidence has been lacking. We are manipulating N form in wet deposition to an ombrotrophic bog, Whim (Scottish Borders), and here report nine years of results. Ammonium and nitrate were provided in rainwater spray as NH4 Cl or NaNO3 at 8, 24 or 56 kg N ha(-1)  yr(-1) , plus a rainwater only control, via an automated system coupled to site meteorology. Detrimental N effects were observed in sensitive nonvascular plant species, with higher cumulative N loads leading to more damage at lower annual doses. Cover responses to N addition, both in relation to form and dose, were species specific and mostly dependent on N dose. Some species were generally indifferent to N form and dose, while others were dose sensitive. Calluna vulgaris showed a preference for higher N doses as ammonium N and Hypnum jutlandicum for nitrate N. However, after 9 years, the magnitude of change from wet deposited N on overall species cover is small, indicating only a slow decline in key species. Nitrogen treatment effects on soil N availability were likewise small and rarely correlated with species cover. Ammonium caused most N accumulation and damage to sensitive species at lower N loads, but toxic effects also occurred with nitrate. However, because different species respond differently to N form, setting of ecosystem level critical loads by N form is challenging. We recommend implementing the lowest value of the critical load range where communities include sensitive nonvascular plants and where ammonium dominates wet deposition chemistry. In the context of parallel assessment at the same site, N treatments for wet deposition showed overall much smaller effects than corresponding inputs of dry deposition as ammonia.

背景与目标： ：氮（N）的湿沉积以氧化（硝酸盐）和还原（铵）形式发生。由于缺乏实地证据，一种形式是否比其他形式更能驱动植被变化。我们正在将N形态以湿沉降的形式运用于营养养生的沼泽Whim（苏格兰边界），这里报告了9年的研究结果。通过耦合到现场气象的自动化系统，在雨水喷雾中以8、24或56 kg N·ha（-1）yr（-1）的铵盐和硝酸盐的形式提供NH4 Cl或NaNO3，加上仅雨水的控制。在敏感的非维管植物物种中观察到有害的氮素影响，较高的累积氮素负荷在较低的年剂量下导致更多的伤害。对氮添加的覆盖响应，无论是形式还是剂量，都是物种特异性的，并且主要取决于氮的剂量。一些物种通常对氮的形式和剂量无动于衷，而另一些则对剂量敏感。寻常的愈伤组织表现出较高的N剂量的偏好，如铵态氮和金丝桃属植物对硝酸盐N的偏好。然而，在9年后，湿法沉积氮在整个物种覆盖率上的变化幅度很小，表明关键物种仅缓慢下降。氮处理对土壤氮素利用率的影响同样很小，很少与物种覆盖率相关。在较低的氮负荷下，铵会引起大部分氮的积累和对敏感物种的破坏，但是硝酸盐也会产生毒性作用。但是，由于不同物种对N形态的反应不同，因此以N形态设定生态系统水平的临界负荷具有挑战性。我们建议在社区包括敏感的非维管植物以及铵在湿法沉积化学中占主导地位的情况下，将临界负荷范围的最小值设为最低值。在同一地点进行并行评估的情况下，与湿法沉积相应的氨投入相比，湿法沉积的N处理总体效果要小得多。