化香果提取物体外抑菌活性研究

研究了化香果提取物对水产常见9种致病菌嗜水气单胞菌（Aeromonas hydrophila）、温和气单胞菌（Aeromonas sobria）、豚鼠气单胞菌（Aeromonas caviae）、维氏气单胞菌（Aeromonas veronii）、迟钝爱德华菌（Edwardsiella tarda）、副溶血弧菌（Vibrio parahaemolyticus）、鼠疫耶尔森氏菌（Yersinia pestis）、大肠杆菌（Escherichia coli）和金黄色葡萄球菌（Staphylococcus aureus）的抑菌活性。研究结果表明：化香果提取物对9种细菌均具有抑制作用,最小抑菌浓度为4.50 g/L。对每种细菌而言,抑菌效果随化香果提取物质量浓度的增加而增强,其中对大肠杆菌的抑制能力最强,IC₅₀值为0.035 g/L;金黄色葡萄球菌次之,IC₅₀值为0.053 g/L。对雌雄小鼠急性经口毒性试验结果表明化香果提取物对小鼠的急性经口半数致死量（LD₅₀值）分别为5 010和7 940 mg/kg体质量,表明化香果提取物属于实际无毒级。     

柔嫩艾美耳球虫诱导鸡巨噬细胞Toll样受体mRNA转录水平的变化

目的探讨柔嫩艾美耳球虫(E.tenella)诱导鸡巨噬细胞Toll样受体(Toll-like receptors,TLRs)m RNA转录水平的变化。方法用有病毒和无病毒E.tenella分别刺激鸡巨噬细胞,于刺激后0、1、2、4、6、8 h收集鸡巨噬细胞,Trizol试剂提取总RNA,经q RT-PCR法检测鸡Toll样受体(chicken TLRs,Ch TLRs)表达的动态变化。结果有病毒E.tenella刺激鸡巨噬细胞的Ch TLR1和Ch TLR21 m RNA的转录水平于刺激4 h时达最高,Ch TLR4、Ch TLR7、Ch TLR15 m RNA的转录水平于刺激2 h时达最高,Ch TLR2、Ch TLR3、Ch TLR5 m RNA转录水平于刺激4 h后显著下调,与对照组比较,差异均有统计学意义(P0.05)。无病毒E.tenella刺激鸡巨噬细胞各时间点的Ch TLR1、Ch TLR4、Ch TLR21 m RNA的转录水平与对照组差异均无统计学意义(P0.05);Ch TLR15 m RNA的转录水平于刺激2 h时达最高,Ch TLR2、Ch TLR3、Ch TLR5、Ch TLR7 m RNA转录水平于刺激4 h后显著下调,与对照组比较,差异均有统计学意义(P0.05)。结论 Ch TLR7和Ch TLR21可能与鸡的抗E.tenella病毒的先天性免疫应答有关,为E.tenella感染的天然免疫机制的研究及鸡E.tenella病的防控奠定了基础。     

青蒿提取物的抑菌性能及在牙膏中的应用

通过最小抑菌浓度（MIC）方法,分析研究和验证青蒿提取物对大肠杆菌、金黄色葡萄球菌、牙龈卟啉单胞菌、变形链球菌的抑菌性能。结果显示：青蒿提取物对以上4种菌具有一定的抑菌活性。采用抑菌率试验方法和二甲苯致小白鼠耳廓急性炎症肿胀模型药理试验方法,试验含青蒿提取物牙膏对大肠杆菌、金黄色葡萄球菌、牙龈卟啉单胞菌、变形链球菌的抗菌功效和抗炎作用。     

对氟苯甲醛缩乙醇胺席夫碱抑菌活性研究

研究了新合成的化合物对氟苯甲醛缩乙醇胺席夫碱对枯草杆菌等五种细菌的抑菌作用。抑菌活性实验表明该席夫碱对枯草杆菌、大肠杆菌、大肠杆菌(101)、金黄色葡萄球菌、革兰氏阴性细菌(发荧光Q67)等5种细菌均有较好的抑菌作用,为寻找具有抑菌、抗癌、抗病毒、杀霉等生物活性的药物提供新途径。     

1H-吲哚亚甲基氨基硫脲席夫碱铜（Ⅱ）配合物及其涂层的抗菌性研究

选用具有生物活性的吲哚-3-甲醛与氨基硫脲制备了席夫碱及其铜（Ⅱ）配合物,通过核磁共振氢谱、红外及紫外光谱等手段鉴定了其结构并确认配合物的配位方式。选用金黄色葡萄球菌（S. aureus）与大肠埃希氏菌（E. coli）测试了席夫碱及其铜（Ⅱ）配合物的抗菌性。结果表明：所制备的铜（Ⅱ）配合物对革兰氏阳性与革兰氏阴性菌均具有较好的抗菌效果,对金黄色葡萄球菌（S. aureus）和大肠埃希氏菌（E. coli）的最小抗菌浓度分别为0.40 g/L和0.95 g/L,具备广谱抗菌性。以吲哚甲醛氨基硫脲席夫碱铜（Ⅱ）配合物作为抗菌剂制备的抗菌涂层同样具有良好的抗菌效果,质量分数为5%时就能够杀灭抗菌涂层上黏附的大肠埃希氏菌（E. coli）。通过电镜观察细菌形貌推测吲哚甲醛氨基硫脲席夫碱铜（Ⅱ）配合物可能通过直接作用于细胞膜将其杀灭,具有较优的抗菌性。     

表皮葡萄球菌发酵提取物对人体皮肤屏障的影响

从健康人体皮肤中分离获取一株表皮葡萄球菌（SE）,并进行培养发酵,制备SE发酵提取物;通过实时无标记动态细胞分析技术（RTCA）检测SE发酵提取物对人原代角质形成细胞（KC）的活性影响;使用聚肌苷酸胞苷酸（poly(I:C)）和脂多糖（LPS）分别刺激KC细胞,造成Toll样受体（TLR）3和TLR 4介导的炎症反应模型,然后加入SE发酵提取物,研究SE发酵提取物对KC细胞炎症反应的抑制作用;最后使用十二烷基硫酸钠（SLS）刺激3D表皮模型,构建受损皮肤模型,并研究SE发酵提取物对损伤皮肤模型中丝聚蛋白（FLG）和兜甲蛋白（LOR）表达影响的作用。结果显示,体积分数为1%SE发酵提取物作用于KC细胞100 h,并未见明显细胞毒性作用,且具有一定的保护作用,SE发酵提取物可以抑制由TLR3介导的炎症反应,而对TLR4介导的炎症反应无明显抑制作用,并且可以有效地提高受损屏障中的FLG和LOR含量。表皮葡萄球菌作为人体皮肤的常驻菌,对皮肤的屏障具有重要的积极作用。     

化妆品防腐剂的短期抑菌效能评价

分别将金黄色葡萄球菌、大肠杆菌的菌悬液与适当浓度的杰马BP、GVL和尼泊金甲酯（以下简称尼甲）3种防腐剂的水溶液或添加防腐剂的化妆品混合均匀,作用4h、8h和24h后进行菌落总数测定,以细菌总数的杀灭对数值作为评价指标。结果显示,尼甲按1：1000-3：1000的比例加入水中后,对大肠杆菌、金黄色葡萄球菌的抑菌作用弱。浓度提高到5：1000～15：1000加入化妆品中后,对大肠杆菌的抑菌效果明硅增强．但对金黄色葡萄球菌的抑杀作用反而减弱。在膏状化妆品中尼甲对大肠杆菌的抑杀作用明显强于液状化妆品。GPL和杰马BP在0．05％～0．15％的水溶液中对2种致病菌产牛明显甚至强烈的抑杀作用,且对2种菌的抑杀强度基本相同,浓度提高到O．25％～0．75％并加入化妆品中后,抑菌效果明显下降。在膏状化妆品中GPL和杰马BP对大肠杆菌的抑杀作用强于金黄色葡萄球菌。膏状和液状化妆品中GPL和杰马BP对大肠杆菌的抑杀作用基本相当。     

乙型流感病毒感染过程中干扰素介导的天然免疫应答

目的 探讨乙型流感病毒(influenza B virus,IBV)感染引起干扰素(interferon,IFN)介导的天然免疫应答。方法以IBV感染犬肾上皮细胞(MDCK)为模型，通过荧光定量PCR法检测IFN信号通路的激活以及IFN刺激基因的表达。收集IBV感染MDCK细胞36及48 h上清，与新鲜培养基混合培养IBV感染MDCK细胞，qPCR法检测内源性IFN的抗病毒作用。加入JAK-STAT通路抑制剂CP后，收集IBV感染MDCK细胞上清，培养IBV感染MDCK细胞，通过qPCR法检测JAK-STAT通路抑制后对内源性IFN抗病毒作用的影响。结果 IBV可有效激活IFN信号通路，并诱导产生Ⅰ型IFN(IFNα、IFNβ)以及Ⅲ型IFN(IFNλ1、IFNλ3)为主的细胞因子。同时，IBV感染MDCK细胞后可诱导产生一系列具有广谱抗病毒作用的IFN刺激基因(IFN-stimulated genes,ISGs)，如ISG15、CCL5、CXCL10、MX1、RIG-I。用JAKSTAT通路抑制剂CP抑制该信号通路后，IBV感染MDCK细胞所诱导生成的ISGs的能力及其相应的抗病毒作...     

A2/O厌氧池污泥同步反硝化聚磷菌增殖、特性诱导及单菌株研究

研究了A2/O厌氧池中具有同步反硝化聚磷功能菌的增殖及诱导前后种群和功能变化。结果表明:经过增殖,设备中反硝化聚磷菌的数量由4.1×103个/mL增加为2.7×106个/mL,磷酸盐的去除率也由38.5%提高到95.1%;诱导后分离到4株假单胞菌属、2株肠杆菌科、1株气单胞菌属、1株葡萄球菌属和1株土壤杆菌属;假单胞菌属、肠杆菌科和葡萄球菌属都是DPB菌,但假单胞菌属的反硝化聚磷功能最强,肠杆菌科的聚磷功能较强,而葡萄球菌属最弱,气单胞菌属和土壤杆菌属既是聚磷菌又是反硝化菌,但不是DPB菌;作为DPB菌应同步具有硝酸盐还原和聚磷的双重生化特性。     

Toll样受体3配体poly(I:C)干预对糖尿病大鼠视网膜病变的影响

目的探讨Toll样受体3(Toll-like receptor 3,TLR3)配体poly(I:C)干预对糖尿病(diabetes mellitus,DM)大鼠视网膜病变的影响。方法用链脲佐菌素(streptozotocin,STZ)建立SD大鼠DM模型,4周后处死大鼠,收集视网膜,Real-time PCR法检测早期DM大鼠视网膜中TLR3和炎性因子(IL-6、IL-1α、TNF-α)mRNA的转录水平,Western blot检测大鼠视网膜中TLR3蛋白的表达水平。分别经正常和4周病程的DM大鼠玻璃体腔注射poly(I:C),48 h后处死大鼠,Real-time PCR检测大鼠视网膜中TLR3和IL-6、IL-1α、TNF-α基因mRNA的转录水平,Western blot检测TLR3蛋白的表达水平,HE染色观察大鼠视网膜结构的变化。结果早期DM组大鼠视网膜中TLR3基因mRNA的转录水平和蛋白的表达水平均显著高于NC组(P均0.01),IL-6、IL-1α、TNF-α基因mRNA的转录水平均显著高于NC组(P均0.01);经玻璃体腔注射poly(I:C),能显著上调大鼠视网膜中TLR3基因mRNA的转录水平和蛋白的表达水平(P0.05或P0.01)以及IL-6和TNF-α基因mRNA的转录水平(P0.05);病程4周时,DM组大鼠视网膜结构与正常大鼠比较,无明显异常,而DM干预组较正常干预组视网膜水肿显著加重,结构出现紊乱。结论 DM大鼠早期视网膜中TLR3的表达显著增加;经玻璃体腔注射poly(I:C)可加重正常和DM大鼠视网膜损伤,可能是通过激活TLR3信号通路,上调信号通路下游产物IL-6、IL-1α、TNF-α的表达水平实现的。     

Expression of Main Toll-Like Receptors in Patients with Different Types of Colorectal Polyps and Their Relationship with Gut Microbiota

Abnormal activation of Toll-like receptor (TLRs) signaling can result in colon cancer development. The aim of this study was to investigate the expression of important TLRs in different histological types of colorectal polyps and evaluate their relationship with intestinal microbiota. The expression levels of TLR2, 3, 4, and 5 were analyzed in intestinal biopsy specimens of 21 hyperplastic polyp (HP), 16 sessile serrated adenoma (SSA), 29 tubular adenoma (TA), 21 villous/tubulovillous (VP/TVP) cases, and 31 normal controls. In addition, selected gut bacteria including Streptococcus bovis, Enterococcus faecalis, Enterotoxigenic Bacteroides fragilis (ETBF), Fusobacterium nucleatum, Porphyromonas spp., Lactobacillus spp., Roseburia spp., and Bifidobacterium spp. were quantified in fecal samples using absolute qRT PCR, and, finally, the association between TLRs and these gut microbiota- was evaluated by Spearman’s correlation coefficient. Higher expression of TLR2 and TLR4 in VP/TVP and TA, and lower expression levels of TLR3 and TLR5 in all type of polyps were observed. The differences in TLR expression patterns was not only dependent on the histology, location, size, and dysplasia grade of polyps but also related to the intestinal microbiota patterns. TLR2 and TLR4 expression was directly associated with the F. nucleatum, E. faecalis, S. bovis, Porphyromonas, and inversely to Bifidobacterium, Lactobacillus, and Roseburia quantity. Furthermore, TLR3 and TLR5 expression was directly associated with Bifidobacterium, Roseburia, and Lactobacillus quantity. Our results suggest a possible critical role of TLRs during colorectal polyp progression. An abnormal regulation of TLRs in relation to gut microbial quantity may contribute to carcinogenesis.     

Distinctive Toll-like Receptors Gene Expression and Glial Response in Different Brain Regions of Natural Scrapie

Prion diseases are chronic and fatal neurodegenerative diseases characterized by the accumulation of disease-specific prion protein (PrP^Sc), spongiform changes, neuronal loss, and gliosis. Growing evidence shows that the neuroinflammatory response is a key component of prion diseases and contributes to neurodegeneration. Toll-like receptors (TLRs) have been proposed as important mediators of innate immune responses triggered in the central nervous system in other human neurodegenerative diseases, including Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis. However, little is known about the role of TLRs in prion diseases, and their involvement in the neuropathology of natural scrapie has not been studied. We assessed the gene expression of ovine TLRs in four anatomically distinct brain regions in natural scrapie-infected sheep and evaluated the possible correlations between gene expression and the pathological hallmarks of prion disease. We observed significant changes in TLR expression in scrapie-infected sheep that correlate with the degree of spongiosis, PrP^Sc deposition, and gliosis in each of the regions studied. Remarkably, TLR4 was the only gene upregulated in all regions, regardless of the severity of neuropathology. In the hippocampus, we observed milder neuropathology associated with a distinct TLR gene expression profile and the presence of a peculiar microglial morphology, called rod microglia, described here for the first time in the brain of scrapie-infected sheep. The concurrence of these features suggests partial neuroprotection of the hippocampus. Finally, a comparison of the findings in naturallyinfected sheep versus an ovinized mouse model (tg338 mice) revealed distinct patterns of TLRgene expression.     

Toll-like Receptor 7 (TLR7) Is Expressed in Adipocytes and the Pharmacological TLR7 Agonist Imiquimod and Adipocyte-Derived Cell-Free Nucleic Acids (cfDNA) Regulate Adipocyte Function

Endosome-localized Toll-like receptors (TLRs) 3 and 9 are expressed and functionally active in adipocytes. The functionality and role of TLR7 in adipocyte biology and innate immunity of adipose tissue (AT) is poorly characterized. We analyzed TLR7 mRNA and protein expression in murine 3T3-L1 and primary adipocytes, in co-cultures of 3T3-L1 adipocytes with murine J774A.1 monocytes and in human AT. The effects of TLR7 agonists imiquimod (IMQ) and cell-free nucleic acids (cfDNA) on adipokine concentration in cell-culture supernatants and gene expression profile were investigated. We found that TLR7 expression is strongly induced during adipocyte differentiation. TLR7 gene expression in adipocytes and AT stroma-vascular cells (SVC) seems to be independent of TLR9. IMQ downregulates resistin concentration in adipocyte cell-culture supernatants and modulates gene expression of glucose transporter Glut4. Adipocyte-derived cfDNA reduces adiponectin and resistin in cell-culture supernatants and potentially inhibits Glut4 gene expression. The responsiveness of 3T3-L1 adipocytes to imiquimod is preserved in co-culture with J774A.1 monocytes. Obesity-related, adipocyte-derived cfDNA engages adipocytic pattern recognition receptors (PRRs), modulating AT immune and metabolic homeostasis during adipose inflammation.     

邻、间、对双胍基苯甲酸盐酸盐抗菌活性的研究

以邻、间、对氨基苯甲酸为起始原料,在盐酸存在下分别与双氰胺合成了中间体邻、间、对双胍基苯甲酸盐酸盐,分别研究了邻、间、对双胍基苯甲酸盐酸盐对革兰氏菌的抗菌活性。实验表明,邻、间、对双胍基苯甲酸盐酸盐对大肠杆菌、金黄色葡萄球菌都具有抑制作用,抗菌活性随浓度的增加而增强;邻双胍基苯甲酸盐酸盐对大肠杆菌、金黄色葡萄球菌最低抑菌浓度分别为0.064mg.mL-1和0.032mg.mL-1,优于间、对双胍基苯甲酸盐酸盐。     

Toll-Like Receptors: Expression and Roles in Otitis Media

Otitis media is mainly caused by upper respiratory tract infection and eustachian tube dysfunction. If external upper respiratory tract infection is not detected early in the middle ear, or an appropriate immune response does not occur, otitis media can become a chronic state or complications may occur. Therefore, given the important role of Toll-like receptors (TLRs) in the early response to external antigens, we surveyed the role of TLRs in otitis media. To summarize the role of TLR in otitis media, we reviewed articles on the expression of TLRs in acute otitis media (AOM), otitis media with effusion (OME), chronic otitis media (COM) with cholesteatoma, and COM without cholesteatoma. Many studies showed that TLRs 1–10 are expressed in AOM, OME, COM with cholesteatoma, and COM without cholesteatoma. TLR expression in the normal middle ear mucosa is absent or weak, but is increased in inflammatory fluid of AOM, effusion of OME, and granulation tissue and cholesteatoma of COM. In addition, TLRs show increased or decreased expression depending on the presence or absence of bacteria, recurrence of disease, tissue type, and repeated surgery. In conclusion, expression of TLRs is associated with otitis media. Inappropriate TLR expression, or delayed or absent induction, are associated with the occurrence, recurrence, chronicization, and complications of otitis media. Therefore, TLRs are very important in otitis media and closely related to its etiology.     

环保增塑剂柠檬酸三丁酯的催化合成工艺研究

采用活性炭固载对甲苯磺酸催化合成了环保增塑剂柠檬酸三丁酯（TBC）。考察了酸醇比、催化剂用量、反应时间、反应温度等工艺条件的变化对柠檬酸三丁酯合成的影响。实验得到制备柠檬酸三丁酯最佳工艺条件：酸醇比为1∶4.0,催化剂质量浓度为2.2%,催化剂负载量为21.0%,反应温度为120℃,反应时间3.0h。在以上条件下,酯化率可达到98.3%。     

Toll-Like Receptor Signaling Pathways: Novel Therapeutic Targets for Cerebrovascular Disorders

Toll-like receptors (TLRs), a class of pattern recognition proteins, play an integral role in the modulation of systemic inflammatory responses. Cerebrovascular diseases (CVDs) are a group of pathological conditions that temporarily or permanently affect the brain tissue mostly via the decrease of oxygen and glucose supply. TLRs have a critical role in the activation of inflammatory cascades following hypoxic-ischemic events and subsequently contribute to neuroprotective or detrimental effects of CVD-induced neuroinflammation. The TLR signaling pathway and downstream cascades trigger immune responses via the production and release of various inflammatory mediators. The present review describes the modulatory role of the TLR signaling pathway in the inflammatory responses developed following various CVDs and discusses the potential benefits of the modulation of different TLRs in the improvement of functional outcomes after brain ischemia.     

Upregulation of TLRs and IL-6 as a Marker in Human Colorectal Cancer

Toll-like receptors (TLRs) not only form an important part of the innate immune system but also serve to activate the adaptive immune system in response to cancer. Real-time PCR; immunohistochemical stain and Western blotting analyses were performed to clarify molecular alterations in colorectal cancer (CRC) patients. We identified Toll-like receptor 1 (TLR1), TLR2, TLR4 and TLR8 gene expression levels and downstream gene, i.e., interleukin-6 (IL-6), IL-8, interferon-α (IFN-α) and myeloid differentiation primary-response protein-88 (MyD88), expression levels in CRC patients and in cancer cell lines. CRC tissues have higher TLR1, TLR2, TLR4, TLR8, IL-6 and IL-8 gene expression levels than do the normal colon mucosa (p < 0.05). TLR2 expression varied in different cell types (mucosa and lymphocytes). There was no difference in the MyD88 and IFN-α gene expression levels between cancerous and normal colon mucosa. CRC patients had higher levels of IL-6 (p = 0.002) and IL-8 (p = 0.038) expression than healthy volunteers did; and higher IL-6 and IL-8 expression was also found to signify a higher risk of recurrence. CL075 (3M002) treatments can reduce the production of IL-8 in different cancer cell lines. The signaling pathway of TLRs in cancer tissue is different from that in normal cells; and is MyD88-independent. Higher expression levels of TLR1, TLR2, TLR 4 and TLR 8 mRNA were related to upregulation inflammatory cytokines IL-6 and IL-8 gene expression in tissue and to the upregulation of IL-6 in blood. The concentration of IL-6 in serum can be used as an indicator of the possibility of CRC recurrence. Treatment with 3M002 can reduce IL-6 production in vitro and may prevent CRC recurrence. Our findings provide evidence that TLR1, TLR2, TLR4 and TLR8 gene expression induce downstream IL-6 and IL-8 gene expression; detection of these expression levels can serve as a CRC marker.     

Toll-Like Receptor and Accessory Molecule mRNA Expression in Humans and Mice as Well as in Murine Autoimmunity,Transient Inflammation,and Progressive Fibrosis

The cell type-, organ-, and species-specific expression of the Toll-like receptors (TLRs) are well described, but little is known about the respective expression profiles of their accessory molecules. We therefore determined the mRNA expression levels of LBP, MD2, CD36, CD14, granulin, HMGB1, LL37, GRP94, UNC93b1, TRIL, PRAT4A, AP3B1, AEP and the respective TLRs in human and mouse solid organs. Humans and mice displayed significant differences between their respective mRNA expression patterns of these factors. In addition, the expression profiles in transient tissue inflammation upon renal ischemia-reperfusion injury, in spleens and kidneys from mice with lupus-like systemic autoimmunity, and in progressive tissue fibrosis upon unilateral ureteral obstruction were studied. Several TLR co-factors were specifically regulated during the different phases of these disease entities, suggesting a functional involvement in the disease process. Thus, the organ- and species-specific expression patterns need to be considered in the design and interpretation of studies related to TLR-mediated innate immunity, which seems to be involved in the tissue injury phase, in the phase of tissue regeneration, and in progressive tissue remodelling.