Mechanism of p53 downstream effectors p21 and Gadd45 in DNA damage surveillance

Both p21 (WAF1/CIP1) and Gadd45 were activated in a p53-dependent manner in MCF-7 cells after being exposed to ionizing radiation. In order to investigate their roles in DNA damage surveillance, p21~(as)/MCF-7 cells stably transfected by p21 antisense expression plasmid pC-WAF1-AS and Gadd45~(as)/MCF-7 stably transfected by Gadd45 antisense expression plasmid pCMVas45 were established. It was observed that G_1 arrest induced by radiation was significantly reduced in Gadd45~(as)/MCF-7 cells as well as in p21~(as)/MCF-7 cells. Repair of radiation damaged report gene greatly reduced in Gadd45~(as)/MCF-7 and p21~(as)/MCF-7 cells. Apoptosis significantly increased in p21~(as)/MCF-7 after exposure to radiation. These results suggest that both p21 and Gadd45 support cellular survival by taking roles in G_1 arrest and DNA repair, furthermore, p21 protects cells from death by inhibiting apoptosis after exposure to ionizing radiation.     

广西、贵州早石炭世层孔虫

The belemnites described here were collected by two field parties visiting southernTibet respectively in 1960 and 1961 from(1)the Lungma—Kapula area,east of Ziang-zhi and(2)Dingje and Dingri,all of which are situated southwest of Lhasa.Theyconsist of two genera and twelve species with six new species and they may be differen-tiated into two assemblages:the first assemblage of upper Oxfordian—lower Tithonianage with Belemnopsis gerardi(Oppel)as its leading species and the second of early Cre-taceous age with Hibolites subfusiformis(Raspail)as its chief element.The geologic sections which bear the belemnites are given as follows(in descendingorder):     

Preparation of Monoclonal Antibodies against Human Ventricular Myosin Light Chain 1 （HVMLC1） for Functional Studies

Using purified recombinant human ventricular myosin light chain 1 (HVMLC 1) as the antigen,three monoclonal antibodies,designated C8,C9 and B 12,were prepared.Immunoblot experiments demonstratedthat all monoclonal antibodies could react with the ventricular myosin light chain 1 isolated from differentsources,such as human,rat or pig.It was also demonstrated that C8 was directed against the NN part of theN-fragment (amino acid 1-40) of HVMLC1,and both C9 and B12 against the C-fragment (amino acid 99-195).The affinity constants of C8,C9 and B12 were 3.20×10~8,8.60×10~7 and 1.77×10~8 M~(-1),respectively,determined by non-competitive ELISA.The isotype of B12 was determined as lgG2a,whereas the isotype ofboth C8 and C9 were IgG1.In the presence of C9 or B12,the actin-activated Mg~(2 )ATPase activity of myosinwas greatly inhibited,but there was almost no effect on the Mg~(2 )ATPase activity for C8.B12 and C9 alsoinhibited the superprecipitation of porcine cardiac native actomyosin (myosin B) and reconstituted actomyosin,but C8 did not.The results indicate that all three monoclonal antibodies could bind the intact myosin molecule,but B12 and C9 might more easily react with epitopes located in the C-fragment of HVMLC1.The inhibitoryeffects of B 12 and C9 on ATPase activity and superprecipitation assays show that light chain 1,particularlythe C-fragment domain,is involved in the modulation of the actin-activated Mg~(2 )ATPase activity of myosinand,as a consequence,plays an essential role in the interaction of actin and myosin.     

美洲南蛇藤的组织培养与快速繁殖

1 植物名称美洲南蛇藤(Celastrus scandens L.). 2 材料类别幼嫩的顶芽或带腋芽的茎段. 3 培养条件(1)芽诱导培养基:1/3MS+6-BA 1.0mg·L-1(单位下同)+IBA 0.02+3%蔗糖;(2)增殖培养基:MS+6-BA 1.0+IBA 0.2+3%蔗糖;(3)壮苗培养基:MS+6-BA 0.1+IBA 0.02+3%蔗糖;(4)生根培养基:1/2MS+NAA 0.5+1.5%蔗糖.     

New Record of Lycodon liuchengchaoi in Anhui

One juvenile and one adult female wolf snake(Colubridae: Lycodon) were sampled at Yixian and Fuxi, Huangshan, Anhui, China in the summer of 2011 and 2012, respectively. The two specimens were identified as Lycodon liuchengchaoi based on external morphology and molecular data. This is a new reptile record in Anhui Province. In our laboratory, four eggs were laid and three neonates were hatched successfully. This is the first record of the laying and incubation of L. liuchengchaoi eggs. The five specimens were deposited at the Museum of Huangshan University(HUM20140001) and Guangdong Entomological Institute(HB-lcfsp12613, HB-lcfsp-ch1~3).     

南蛇藤的组织培养和快速繁殖

1植物名称南蛇藤(Celastrus orbiculatus Thunb),又名落霜红、霜红藤、南蛇风. 2材料类别幼嫩的顶芽或带腋芽的茎段. 3培养条件(1)起始培养基:1/3MS 6-BA 1.0 mg·L-1(单位下同) IBA 0.02 3%蔗糖;(2)增殖培养基:MS 6-BA 1.2 IBA 0.2 3%蔗糖;(3)生根培养基:1/2 MS NAA 0.3 1.5%蔗糖.上述培养基均加入6.0 g·L-1倍力凝(一种微生物多糖固化剂),pH 5.8～6.2.培养温度为18～26℃,培养室内全自然光照,光照度100～2 000 1x,光照时间9～13 h·d-1.     

Two New Triterpenes from the Orchid Pholidota yunnanensis Rolfe

Two novel triterpenes (1 and 2) were isolated from the orchid Pholidota yunnanensis Rolfe.Using chemical and spectral analyses (UV, IR, MS, 1D-NMR and 2D-NMR), these two triterpenes were established as 25-methylenecyclopholidonyl p-hydroxy-trans-cinnamate (1; named pholidotine A) and 25methylenecyclopholidonyl p-hydroxy-cis-cinnamate (2; named pholidotine B).     

Influence of silicon on microdistribution of mineral ions in roots of salt-stressed barley as associated with salt tolerance in plants

Two contrasting barley (Hordeum vulgare L.) cultivars: Kepin No.7 (salt sensitive), and Jian 4 (salt tolerant) were grown hydroponically to investigate the microdistribution of mineral ions in roots as affected by silicon (Si) with respect to salt tolerance. The experiment was undertaken consisting of two treatments with 3 replicates: (i) 120 mmol · L-1 NaCl alone (referred to as Si-NaCl+), (ii) 120 mmol·L-1 NaCl + 1.0 mmol · L-1 Si (as potassium silicate) (referred to as Si+NaCl+). Plant root tips were harvested for microanalysis using an energy dispersive X-ray microanalyzer (EDX) 30 d after transplanting. Higher Cl and Na X-ray peaks were recorded in the root epidermal, cortical and stelar cells of roots for the treatment Si-NaCl+ with the majorities of Na and Cl being accumulated in epidermal and cortical cells, while relatively low K peaks were observed regardless of the barley cultivars used. By contrast, considerably higher K peaks were detected in the epidermal, cortical and stelar cells of th     

重组腺相关病毒2型转导人外周血单核细胞来源树突状细胞的研究

To find out the infection efficiency of recombinant adeno-as sociated virus 2-mediated exogenou s genes in human peripheral blood monocyte-derived dendritic cells(D Cs),the process of transfection wa s investigated by FITC-labeled rAA V2,and observed under confocal mic roscope.Newly separated dendritic cells were tranfected by rAAV2-luc and rAAV2-GFP at different MOI,and transfection efficiency were detec ted by luminometer for rAAV2-luc a nd flow cytometry for rAAV2-GFP.Th e results were elucidated in four different assay systems:(1)60min w as needed for rAAV2 to bind on den dritic cells,and got into cells in the following 10min;(2)the express ion of luc could be detected at th e MOI as low as 1×10~5v.g/cell,and the expression plateau was reached by the MOI of 10~6～10~7v.g/cell,furt her increase of MOI had no functio n on expression level;(3)transgene expression was detected after 48h, and maintained a higher expression level from 96h to 240h after infec tion;(4)7days postinfection of rAA V2-GFP,5%～18% dendritic cells were GFP positive. These data suggest th at rAAV2 vector can efficiently in fect monocyte-derived dendritic ce lls and mediate exogenous gene exp ression,and that the application o f rAAV2 as vector may be useful fo r gene transfer to dendritic cells in ex vivo immunotherapy.     

毛枝南蛇藤根皮中南蛇藤素的分离鉴定及含量测定

从卫矛科南蛇藤属植物毛枝南蛇藤(Celastrus hookeri Prain)根皮中首次分离出一个红色立方体形结晶,经紫外、红外、高分辨质谱和~(13)C核磁共振谱分析,并与有关文献数据对照,确认是南蛇藤素。作者用薄层扫描和高效液相两种方法初步测定了毛枝南蛇藤根皮和根的木栓化表皮的南蛇藤素含量,结果表明南蛇藤素主要集中在根的木栓化表皮,含量在12%以上,而在整个根皮中的含量大约为1%。     

Structural analysis of DMD gene and its clinical application in Chinese.I.Bgl Ⅱ exon—containing fragment,RFLP and carrier detection

This article is one of the serial studies on the characteristics of the molecular structure for dystrophin gene in Chinese.By using the entire dystrophin cDNA(14kb) as a probe,the number and RFLPs of Bgl Ⅱ exon-containing fragments of the dystrophin gene were analysed.Four new Bgl Ⅱ fragments were found,two of them(3.7 and 6.2 kb) detected by comparing the hybridization patterns with cDNA1-2a,1a and 2a,one(9.3 kb) from the hybridization pattern with cDNA 9 by lengthening migrating distance of DNA fragments in electrophoresis,and another and (4.0 kb) by comparing the patterns with cDNA 11-14, 11a,11b,aac-12a and 14.The results indicated that the number of Bgl Ⅱ exon-containing fragments should be 59 rather than 55 reported previously,which laid the foundation of the Bgl Ⅱ partial restriction map for dystrophin gene.Three of the four RFLPs found in Caucacian appear in the hybridization patterns of three subclones,i.e. cDNA 2b-3,cDNA 4-5,and cDNA 5b-7.The values of expected heterozygote frequency(EHF) were 0.33,0.33 and 0.40,and the observed heterozygote frequency(OHF) were 0.40,0.40 and 0.48 respectively.Meanwhile,two new rare allelic fragments(15kb) were found in RFLPs from Bgl Ⅱ/2b-3 and Bgl Ⅱ/4-5a patterns respectively.These Bgl Ⅱ RFLPs and four XbaI RFLPs documented in our laboratory have been used to detect the carrier in 7 DMD families and 1 BMD family.Of the 69 individuals from the 8 families,11 females were diagnosed as the carriers with DMD mutation,4 females as the doubtful carriers,12 females were defined as normal genotype and 2 females as probably normal.The results suggest that the carrier testing method based on dosage intensity analysis and genotype analysis by using dystrophin cDNA as a probe will be more sensitive and accurate.     

Regulated genes in mesenchymal stem cells and gastric cancer

AIM: To investigate the genes regulated in mesenchymal stem cells(MSCs) and diffuse-type gastric cancer(GC),gene expression was analyzed. METHODS: Gene expression of MSCs and diffuse-type GC cells were analyzed by microarray. Genes related to stem cells, cancer and the epithelial-mesenchymal transition(EMT) were extracted from human gene lists using Gene Ontology and reference information. Gene panels were generated, and messenger RNA gene expression in MSCs and diffuse-type GC cells was analyzed. Cluster analysis was performed using the NCSS software.RESULTS: The gene expression of regulator of G-protein signaling 1(RGS1) was up-regulated in diffuse-type GC cells compared with MSCs. A panel of stem-cell related genes and genes involved in cancer or the EMT were examined. Stem-cell related genes, such as growth arrest-specific 6, musashi RNA-binding protein 2 and hairy and enhancer of split 1(Drosophila), NOTCH family genes and Notch ligands, such as delta-like 1(Drosophila) and Jagged 2, were regulated.CONCLUSION: Expression of RGS1 is up-regulated, and genes related to stem cells and NOTCH signaling are altered in diffuse-type GC compared with MSCs.     

Secretory/releasing proteome-based identification of plasma biomarkers in HBV-associated hepatocellular carcinoma

For successful therapy, hepatocellular carcinoma (HCC) must be detected at an early stage. Herein, we used a proteomic approach to analyze the secretory/releasing proteome of HCC tissues to identify plasma biomarkers. Serum-free conditioned media (CM) were collected from primary cultures of cancerous tissues and surrounding noncancerous tissues. Proteomic analysis of the CM proteins permitted the identification of 1365 proteins. The enriched molecular functions and biological processes of the CM proteins, such as hydrolase activity and catabolic processes, were consistent with the liver being the most important metabolic organ. Moreover, 19% of the proteins were characterized as extracellular or membrane-bound. For validation, secretory proteins involved in transforming growth factor-β signaling pathways were validated in plasma samples. Alphafetoprotein (AFP), metalloproteinase (MMP)1, osteopontin (OPN), and pregnancy-specific beta-1-glycoprotein (PSG)9 were significantly increased in HCC patients. The overall performance of MMP1 and OPN in the diagnosis of HCC remained greater than that of AFP. In addition, this study represents the first report of MMP1 as a biomarker with a higher sensitivity and specificity than AFP. Thus, this study provides a valuable resource of the HCC secretome with the potential to investigate serological biomarkers. MMP1 and OPN could be used as novel biomarkers for the early detection of HCC and to improve the sensitivity of biomarkers compared with AFP.     

Pharmacokinetics of recombinant human interleukin-3 in rhesus monkeys

Concentration-time profiles of ~(125)I-labeled recombinant human interleukin-3 (125IrhIL-3) were de-termined by reverse phase high performance liquid chromatography (RHPLC) after intravenous and subcutaneous ad-ministration of the drug in 16 rhesus monkeys. The initial and terminal T1/2 in plasma after intravenous of 30 μg/kg were (0. 15 ±0.13) and (2. 21 ± 0. 59) h, respectively. Terminal half-lives after 30, 90 and 180μg/kg subcutaneous (s.c.) injections were 2. 0-3.8 h. Area under concentration-time curves (AUC) following s. c. were roughly in-creased with dose, while CL5 were similar among different dosages. The absorption rates were dependent on concentra-tion at injected site. Bioavailability was about 0.7 after s.c. Rapid biodegradation was found in plasma. Distribution profiles of total radioactivity were as follows: the highest level was found in urinary system; levels in bile-enteric sys-tem, lymph nodes, bone marrow and spleen were near to that in plasma, and level in brain was the lowest     

Isolation of ScFv antibodies of rP27~(Kip1) from phage display libraries constructed from immunized and non-immunized repertoires

Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27~(Kipl)immunized and non-immunized mice. After only one round of selection with rP27~(Kipl), clones from each library were chosen randomly and digested by Taq I and Hinf I. 11 of 64 clones from the immunized animal had consistent restriction pattern, while none of the 64 clones from the non-immunized animal had, except that one had the same fragments pattern as that of the 11 clones. The 12 fragments were expressed in E. coli BL21(DE3)/pET-28b( ) system. ELISA showed that some of the fragments could bind to rP27~(Kipl) specifically. All these results implied that specific antibody can be obtained by genetic engineering without hybridoma or many rounds of growth and panning selection.     

Two New Triterpenoids from the Roots of Sanguisorba officinalis L.

Two new triterpenoids, octanordammar-1,11,13(17)-trien-17-ol-3,16-dione (1) and lup-12-en-15α,19β-diol-3,11-dioxo-28-oic acid (4), as well as 13 known compounds were isolated from the roots of Sanguisorba officinalis L. (Rosaceae). Their structures were determined using spectroscopic methods.     

南蛇藤脯氨酸脱氢酶基因的克隆和表达特性

利用兼并性引物和RACE方法, 在南蛇藤(Celastrus orbiculatus)中克隆了1个脯氨酸脱氢酶基因, 并命名为NstProDH1。序列比对显示该基因与拟南芥(Arabidopsis thaliana)和烟草(Nicotiana tabacum)的脯氨酸脱氢酶(ProDH)具有很高的同源性。酶学特性分析表明该酶具有脯氨酸脱氢酶的活性。比较南蛇藤不同器官该基因的转录表达模式与脯氨酸脱氢酶活性, 结果显示两者之间没有明显的关联, 说明该基因的表达受到转录和翻译水平的双重调控, 同时也暗示南蛇藤中还存在其它的脯氨酸脱氢酶基因。NstProDH1基因的表达模式与野生型拟南芥中的ProDH1具有相似性, 因此推测NstProDH1基因可能在功能上与拟南芥ProDH1基因相似。     

螺旋藻品系Z中不同形态藻丝体的生长速率和光合色素比较

The line-shaped filaments Sp-Z(L) were isolated and cultured from Spirulina platensis Sp-Z. The growth rate of Sp-Z(L) was only 64% as much as that of SP-Z when the light intensity was 4000 Lux. The contentS (×10-3 g / g dry weight) of chlorophylls, carotenoids and phycobilins of Sp-Z(L) and Sp-Z were 20.6, 0.343, 5.00 and 24.1 0.297, 4.46, respectively. Moreover, as to the absorption spectra of the three photopigments of SP-Z(L), red shifts were observed. Therefore, after the spiral Sp-Z breeded or changed…     

Cell-specific regulation of APOBEC3F by interferons

Human cytidine deaminase APOBEC3F(A3F)has broad anti-viral activity against hepatitis Bvirus and retroviruses including human immunodeficiency virus type 1.However,its regulation in viralnatural target cells such CD4~ T lymphocytes,macrophages,and primary liver cells has not been wellstudied.Here we showed that A3F was up-regulated by interferon(IFN)-α in primary hepatocytes andmultiple liver cell lines as well as macrophages.Although the IFN-α signaling pathway was active in Tlymphoid cells and induction of other IFN stimulated genes such as PKR was detected,A3F and APOBEC3G(A3G)were not induced by IFN-α in these cells.Thus,additional factors other than known IFN-stimulatedgenes also regulated IFN-α-induced A3F expression distinctly.A3F and A3G expression levels in primaryhepatocytes,especially after IFN-α stimulation,were comparable to those in CD4~ T lymphocytes in someindividuals.Significant variations of A3F and A3G expression in primary hepatocytes from various subjectswere observed.Individual variations in A3F and/or A3G regulation and expression might influence the clinicaloutcomes of hepatitis B infection.     

Molecular and in vitro Characterization of Field Isolates of Bovine Herpesvirus-1

Bovine Herpesvirus-1 (BoHV-1) is distributed worldwide and is a major pathogen in cattle, being the causal agent of a variety of clinical syndromes. The aim of this study was to isolate and to characterize (molecular and biological characterization) BoHV-1 from 29 immunosuppressed animals. It was possible to obtain 18 isolates, each from a different animal, such as from the respiratory and reproductive tracts. In some cases the cytopathic effect was visible 12 hours post-inoculation, and became characteristic after 36-48 hours. Biological characteristics were evaluated and compared with Iowa and Colorado-1 reference strains, and differences were found in plaque size, virus titer measured by TCID50 and PFU/mL, and one step virus curves. These results showed that some isolates had a highly virulent-like behavior in vitro, compared to the reference strains, with shorter eclipse periods, faster release of virus into the supernatants, and higher burst size and viral titer. There were no differences in glycoprotein expression of BoHV-1 isolates, measured by Western blot on monolayers. Moreover, using restriction endonucleases analysis, most of the viruses were confirmed as BoHV-1.1 and just one of them was confirmed as BoHV-1.2a subtype. These findings suggest that some wild-type BoHV-1 isolates could be useful as seeds to develop new monovalent vaccines.