复方甘草酸苷片与他克莫司软膏在湿疹和特应性皮炎治疗中的应用

目的：分析复方甘草酸苷片与他克莫司软膏在湿疹和特应性皮炎治疗中的应用效果，为临床工作提供有价值参考。方法：收集分析2015年5月～2016年5月我院收治的湿疹和特应性皮炎患者50例作为研究对象，随机分为实验组和对照组，对照组采用糠酸莫米松乳膏治疗，实验组使用复方甘草酸苷片联合他克莫司软膏，比较两组临床效果。结果：实验组治疗总有效率92.0%，明显高于对照组68.0%，对照组不良反应总发生率44.0%，明显高于实验组20.0%，差异显著，具有统计学意义（P＜0.05）。结论：在湿疹和特应性皮炎的治疗中使用复方甘草酸苷片和他克莫司软膏，具有良好的临床效果，疗效确切、安全可靠，不良反应少。

A MONOCLONAL ANTIBODY RECOGNIZING NON DERIVATIVE 13 HYDROXY GIBBERELLINS AND THEIR GLUCOSIDES *

The production and characterization of a monoclonal antibody (MAb AB10) against GA 3 glucoside as well as GA 3 is described. MAb AB10 was derived from an immunogen in which human serum albumin (HSA) was linked to GA 3 at carbon 3. This antibody showed high affinity for GA 3 glucoside as well as for 13 hydroxy gibberellins (GA 1, GA 3, GA 5, etc). The affinity of MAb AB10 for 13 hydroxy GAs was significantly reduced by methylation of the 7 oic acid but not by glycosylation of 3 hydroxyl group. Based on this antibody, both of competitive enzyme linked immunosorbent assays (ELISAs) for GA 3 glucoside and for GA 3 were developed. These two ELISAs displayed linear detection ranges from 0 2 pmol to 20 pmol. Using these assays, the fluctuation of GA 3 like and GA 3 glucoside like substances in the leaves of Rumex japonicus was investigated. The results indicated that the glycosylation of free GAs was connected with leaf senescence and that the function of 6 benzyl amino purine in retarding the leaf senescence was probably related to delaying the process of glycosylation of free GAs.

Outcomes of CAG Regimen for Refractory Biphenotypic Acute Leukemia Patients

Objective To evaluated the efficiency of low-dose cytosine arabinoside plus aclarubicin with concurrent administration of granulocyte colony-stimulating factor （CAG） regimen for refractory biphenotypic acute leukemia （BAL）. Methods We treated 5 refractory BAL patients by CAG regimen （10 mg.m 2 cytosine arabinoside subcutaneously administrated every 12 hours, day 1-14; 5-7 mg·m^-2 aclarubicin intravenously administrated daily, day 1-8; and concurrently used 200 μg·m^-2·d^-1 granulocyte colony-stimulating factor subcutaneously） from November 2002 to April 2007. The efficacy of the regimen was evaluated by response rate, and the side effects were also measured. Results The complete remission rate was 80%, median duration of absolute neutrophil count〈5.0×10^8/L and platelet count〈2.0×10^10/L was day 13 and day 1, respectively; and the infection rate was low （Ⅲ-Ⅳ infection rate, 20.00%）.

Influence of a nucleotide oligomerization domain 1 (NOD1) polymorphism and NOD2 mutant alleles on Crohn's disease phenotype

AIM： To examine genetic variation of nucleotide oligomerization domain 1 （NOD/） and NOD2, their respective influences on Crohn＇s disease phenotype and gene-gene interactions. METHODS： （ND1＋32656＊1） NOD1 polymorphism and SNPS, SNP12 and SNP13 of NOD2 were analyzed in 97 patients and 50 controls. NOD2 variants were determined by reaction restriction fragment length polymorphism analysis. NOD1 genotyping and NOD2 variant confirmation were performed by specific amplification and sequencing. RESULTS： The distribution of NOD1 polymorphism in patients was different from controls （P = 0.045） and not altered by existence of NOD2 mutations. In this cohort, 30.92% patients and 6% controls carried at least one NOD2 variant （P 〈 0.001） with R702W being the most frequent variant. Presence of at least one NOD2 mutation was inversely associated with colon involvement （9.09% with colon vs 36.4% with ileal or ileocolonic involvement, P = 0.04） and indicative of risk of penetrating disease （52.63% with penetrating vs 25.64% with non-penetrating or stricturing behavior, P = 0.02）. L1007finsC and double NOD2 mutation conferred the highest risk for severity of disease （26.3% with penetrating disease vs 3.8% with non-penetrating or stricturing behavior presented L1007finsC, P = 0.01 and 21.0% with penetrating disease vs 2.5% with non-penentrating or stricturing behavior carried double NOD2 mutation, P = 0.007）. Exclusion of patients with NOD2 mutations from phenotype/NODl-genotype analysis revealed higher prevalence of ＊1＊1 genotype in groups of younger age at onset and colonic location.CONCLUSION： This study suggests population differences in the inheritance of risk NOD1 polymorphism and NOD2 mutations. Although no interaction between NOD1-NOD2 was noticed, a relationship between disease location and Nod-like receptor molecules was established.

Heterozygous nucleotide-binding oligomerization domain-2 mutations affect monocyte maturation in Crohn's disease

AIM： To investigate the function of monocytes in Crohn＇s disease （CD） patients and to correlate this with disease- associated nucleotide-binding oligomerization domain-2 （NOD2） gene variants. METHODS： Monocytes from 47 consecutively referred CD patients and 9 healthy blood donors were cultured with interleukin （IL）-4 and granulocyte-macrophage colony-stimulating factor （GM-CSF）, and stimulated with lipopolysaccharide （LPS） or muramyldipeptide （MDP）, the putative ligand of NOD2. RESULTS： We found that monocytes from CD patients differentiated in vitro to mature dendritic cells （DCs）, as determined by immunophenotype and morphology. IVOD2 genotype was assessed in all subjects, and we observed high CD86 expression on immature and LPS-stimulated DCs in IVOD2 mutated CD patients, as compared with wtlVOD2 CD patients and controls. By contrast, CD86 expression levels of DCs induced to maturity with MDP derived from IVOD2-mutated subjects were comparable to those of normal subjects. The amount of IL-12p70 in patient-cell cultures was larger than in controls aEer LPS treatment, but not aEer treatment with MDR CONCLUSION： Our results suggest that DCs obtained from patients with mutations in the IVOD2 gene display an activated phenotype characterized by high CD86 expression, but have a diminished response to MDP when compared to the terminal differentiation phase. We speculate that the altered differentiation of monocytes might lead to an imbalance between inflammation and the killing ability of monocytes, and may be relevant to the pathogenesis of CD.

Heat shock protein 70 antisense oligonucleotide inhibits cell growth and induces apoptosis in human gastric cancer cell line SGC-7901

AIM: Heat shock protein (HSP)70 is over-expressed in human gastric cancer and plays an important role in the progression of this cancer. We investigated the effects of antisense HSP70 oligomer on human gastric cancer cell line SGC-7901, and its potential role in gene therapy for this cancer.METHODS: Human gastric cancer cell line SGC-7901 was treated in vitro with various concentrations of antisense HSP70 oligonucleotides at different intervals. Growth inhibition was determined as percentage by trypan blue dye exclusion test. Extracted DNA was electrophoresed on agarose gel, and distribution of cell cycle and kinetics of apoptosis induction were analyzed by propidium iodide DNA incorporation using flow cytometry, which was also used to detect the effects of antisense oligomer pretreatment on the subsequent apoptosis induced by heat shock in SGC-7901 cells. Proteins were extracted for simultaneous measurement of HSP70 expression level by SDS-PAGE Western blotting.RESULTS: The number of viable cells decreased in a doseand time-dependent manner, and ladder-like patterns of DNA fragments were observed in SGC-7901 cells treated with antisense HSP70 oligomers at a concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h, which were consistent with inter-nucleosomal DNA fragmentation. Flow cytometric analysis showed a dose- and time-dependent increase in apoptotic rate by HSP70 antisense oligomers. This response was accompanied with a decrease in the percentage of cells in the G1 and S phases of the cell cycle, suggesting inhibition of cell proliferation. In addition, flow cytometry also showed that pretreatment of SGC-7901 cells with HSP70 antisense oligomers enhanced the subsequent apoptosis induced by heat shock treatment. Western blotting demonstrated that HSP70 antisense oligomers inhibited HSP70 expression, which preceded apoptosis, and HSP70 was undetectable at the concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h.CONCLUSION: Antisense HSP70 oligomers can abrogate HSP70 expression in SGC-7901 cells, which may in turn induce apoptosis and inhibit cell proliferation, conversely suggesting that HSP70 is required for the proliferation and survival of human gastric cancer cells under normal conditions.

Cloning and Sequence Analysis of Envelope Glycoprotein E1 Gene of Rubella Virus, JR23 Strain

To construct an expression vector containing the E1 glycoprotein gene of rubella virus for the study on the effect of mutation of the E1 gene glycoprotein and the analysis of phylogenetic differences of sequences, the gene encoding the E1 envelope glycoprotein was amplified from rubella virus, Jinan strain JR23, by RT-PCR and ligated into PMD-18T vector. The clones that carried the E1 gene were identified after amp r selection and analysis of restriction enzyme digestion. After sequencing this gene was analyzed by Danstar and Winstar programs, and the map of phylogenetic tree was drawn. The clone of E1 glycoprotein was thus constructed. It was found that the sequence differences between JR23 strain and the TCRB strain from Japan and those between JR23 strain and Thomas strain of England were rather small with difference values of 0.9% and 1.2% respectively. Yet those between JR23 strain and BRD2 strain from Beijing and those between JR23 strain and XG379 strain from Hong Kong were comparatively larger with difference values of 7.6% and 7.3% respectively. The sequence of JR23 strain with other strains was less than 3% except the NC strain (3.7%). It concludes that the construction of E1 glycoprotein gene offers an approach to study the relationship between structures and functions of E1 gene and its gene products. In the phylogenetic tree, it shows that there are significant differences in the sequences of rubella virus isolated in China, and this might be helpful to develop an effective subunit vaccine.

高效毛细管电泳法拆分25R、25S鲁斯可皂苷元初步研究

目的:对高效毛细管电泳法拆分25R、25S-鲁斯可皂苷元混合物的条件进行探讨,以建立一种快速测定鲁斯可皂苷元差向异构体比例的方法。方法:以羟丙基-β-环糊精作为手性添加剂,考察分离电压、缓冲液pH、离子浓度、环糊精浓度对25R、25S-鲁斯可皂苷元拆分的影响。结果:从湖北麦冬制备分离的鲁斯可皂苷元,NMR、IR检测结果显示其是以25 S构型为主要成分、25R和25S 2个差向异构体并存的混合物。以0.018 mol·L-1的羟丙基-β-环糊精为手性添加剂,分离电压6.0 kV,0.075 mol·L-1的pH=10.0磷酸氢二钠缓冲液,25R与25S-鲁斯可皂苷元可实现基线分离(分离度1.52±0.13),在此条件下,测得制备的鲁斯可皂苷元中25 R、25 S异构体含量比为25.4∶74.6。结论:高效毛细管电泳法可用于25R与25S-鲁斯克皂苷元手性拆分。研究结果为鲁斯可皂苷元药理学研究、鲁斯可皂苷元质量控制提供新方法。

Trans-generational effects in the clonal invader Alternanthera philoxeroides

Aims Recent studies have revealed heritable phenotypic plasticity through vegetative generations.In this sense,changes in gene regulation induced by the environment,such as DNA methylation(i.e.epigenetic changes),can result in reversible plastic responses being transferred to the offspring generations.This trans-generational plasticity is expected to be especially relevant in clonal plants,since reduction of sexual reproduction can decrease the potential for adaptation through genetic variation.Many of the most aggressive plant invaders are clonal,and clonality has been suggested as key to explain plant invasiveness.Here we aim to determine whether trans-generational effects occur in the clonal invader Alternanthera philoxeroides,and whether such effects differ between populations from native and non-native ranges.Methods In a common garden experiment,parent plants of A.philoxeroides from populations collected in Brazil(native range)and Iberian Peninsula(non-native range)were grown in high and low soil nutrient conditions,and offspring plants were transplanted to control conditions with high nutrients.To test the potential role of DNA methylation on trans-generational plasticity,half of the parent plants were treated with the demethylating agent,5-azacytidine.Important Findings Trans-generational effects were observed both in populations from the native and the non-native ranges.Interestingly,trans-generational effects occurred on growth variables(number of ramets,stem mass,root mass and total mass)in the population from the native range,but on biomass partitioning in the population from the non-native range.Trans-generational effects of the population from the native range may be explained by a‘silver-spoon’effect,whereas those of the population from the non-native range could be explained by epigenetic transmission due to DNA methylation.Our study highlights the importance of trans-generational effects on the growth of a clonal plant,which could help to understand the mechanisms underlying expansion success of many clonal plants.

Molecular characterization and infectivity of Papaya leaf curl China virus infecting tomato in China

Papaya leaf curl China virus (PaLCuCNV) was previously reported as a distinct begomovirus infecting papaya in southern China. Based on molecular diagnostic survey, 13 PaLCuCNV isolates were obtained from tomato plants showing leaf curl symptoms in Henan and Guangxi Provinces of China. Complete nucleotide sequences of 5 representative isolates (AJ558116, AJ558117, AJ704604, FN256260, and FN297834) were determined to be 2738–2751 nucleotides, which share 91.7%–97.9% sequence identities with PaLCuCNV isolate G2 (AJ558123). DNA-β was not found to be associated with PaLCuCNV isolates. To investigate the infectivity of PaLCuCNV, an in-fectious clone of PaLCuCNV-[CN:HeNZM1] was constructed and agro-inoculated into Nicotiana benthamiana, N. tabacum Samsun, N. glutinosa, Solanum lycopersicum and Petunia hybrida plants, which induced severe leaf curling and crinkling symptoms in these plants. Southern blot analysis and polymerase chain reaction (PCR) indicated a systemic infection of test plants by the agro-infectious clone.

Study on the regulatory effect of herbal cake-partitioned moxibustion on colonic CD206,AMPK and TSC2 in rats with Crohn disease

Objective:To explore the mechanism of herbal cake-partitioned moxibustion in Crohn disease(CD)treatment by observing the effect of herbal cake-partitioned moxibustion on protein expressions of colonic M2 macrophage marker CD206,AMP-activated protein kinase(AMPK)and tuberous sclerosis complex(TSC)2.Methods:Twenty-six specific pathogen free male rats were randomly divided into a normal group,a model group and a herbal cake-partitioned moxibustion group.The CD model was prepared by enema with the mixture of 5%(W/V)2,4,6-trinitrobenzene sulfonic acid(TNBS)and 50%ethanol at 2:1(volume ratio).After the model was successfully prepared,rats in the herbal cake-partitioned moxibustion group received herbal cake-partitioned moxibustion at Qihai(CV 6)and bilateral Tianshu(ST 25).Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of rat colon;immunohistochemical technique was used to detect the expression of colonic CD206 protein;Western blot,immuno fluorescence,and real-time fluorescence quantitative polymerase chain reaction(RTqPCR)technologies were used to detect the protein and mRNA expressions of colonic AMPK and TSC2.Results:Compared with the normal group,rats in the model group showed damaged colonic mucosa,missing of the epithelial layer;thickened submucosa,vascular proliferation,massive infiltration of monocytes and lymphocytes,and cracked ulcers that reached the muscle layer.Rats in the herbal cake-partitioned moxibustion group showed reduced intestinal inflammation and healing intestinal epithelium ulcers.Compared with the normal group,rat colonic CD206 protein expression,and the protein and mRNA expressions of colonic AMPK and TSC2 were decreased in the model group(all P<0.Ol);compared with the model group,rat colonic CD206 protei n expression was in creased(P<Q.Q1),as well as the protein and mRNA expressions of AMPK and TSC2 in the herbal cake-partitioned moxibustion(all P<0.05).Conclusion:Herbal cake-partitioned moxibustion can reduce intestinal inflammation in CD rats,increase colonic CD206 protein expression,and up-regulate the protein and mRNA expressions of colonic AMPK and TSC2.