Etiological analysis of clinical cases of live attenuated varicella vaccine / 中国生物制品学杂志

@#Objective To analyze the etiology of clinical cases of live attenuated varicella vaccine.Methods 64 samples of varicella vesicle fluid from 49 patients clinically diagnosed as varicella cases in phase Ⅲ clinical trial of live attenuated varicella vaccine in enterprises(the test site was Henan Province) were collected,extracted for DNA,and distinguished for wild-type strains and vaccine strains by PCR and restriction fragment length polymorphism(PCR-RFLP).Genotype was analyzed using the genotyping method recommended at the international(varicella-zoster virus,VZV) nomenclature meeting2008.Results 64 vesicle fluids were all wild-type strains(Pst Ⅰ~+Sma Ⅰ~-BssH Ⅱ~-Nae Ⅰ~-),and no vaccine-related cases occurred.All 49 isolates belonged to Clade 2.Additionally,compared with Clade 2,a synonymous mutation(T→C) in SNP18 082 was detected in all 49 isolates,and a mutation(C→A) in SNP790 was detected in one isolate.Conclusion In the clinical trial of live attenuated varicella vaccine in Henan Province,all the clinical cases were caused by infection of wild-type strain which belonged to Clade 2 genetic branch.

Phenotypic characteristics of wild-type plague phage growth in different experimental environments / 中国热带医学

@#Abstract: Objective To observe the phenotypic characteristics of 3 wild-type plague phages under different experimental environments, providing scientific evidence for the identification of phage biological characteristics and the study of their interaction with host bacteria in the future. Methods The sensitivity of 3 wild-type plague phages were detected by using liquid culture method, emisolid medium method and micro-liquid culture method based on OmniLog TM microbial identification system. Results The growth result based on LB liquid medium showed that the growth of plague phage 476 for 20-24 hours at both 28 ℃ and 37 ℃was better than that of plague phages 087 and 072204 at 37 ℃, and the growth of plague phages 087 was better than that of plague phages 072204 at 37 ℃. With the attenuated plague bacterium EV76 as the host bacterium, phage 476 was able to form visible plaque on double-layer agar medium for 20-20 hours at both 28 ℃ and 37 ℃, phages 087 and 072204 were only able to form opaque plaque on double-layer agar medium for 20-24 hours at 37 ℃. The growth results based on OmniLogTM system showed that when plague phage was lysed in EV76 strain at 33 ℃, the first row appeared as a straight line with a peak of no more than 100 in the 96-well microplate curve chart. As the phage quantity decreased, the dilution plate appeared with growth curve similar to EV76 strain in turn, and the color of tetrazolium dyes in the experimental wells gradually deepened as the phage number decreased and the host bacteria number increased. Therefore, it indicates that phage 476 was sensitively at both 28 ℃ and 37 ℃, while phage 087 and 072204 were temperature-dependent only at 37 ℃ to attenuated plague bacterium EV76. Conclusions The lysing ability of 3 wild-type plague phages are temperature-dependent, and the growth results are consistent under the three experimental conditions.

Study of neurotrophic factor receptor tyrosine kinase gene fusion in the precise treatment of wild-type gastrointestinal stromal tumor / 中华胃肠外科杂志

The neurotrophin receptor kinase (NTRK) gene encodes neurotrophic factor receptor tyrosine kinase (NTRK), which plays an important role in the development and function of the nervous system. NTRK gene fusion mutation results in the production of chimeric NTRK proteins, which have carcinogenic potential through constitutive activation or overexpression. NTRK gene fusion mutation can lead to a special type of wild type gastrointestinal stromal tumor (GIST), whose clinical manifestations and treatment are completely different from other types of GIST. This fusion mutation can be detected clinically by a variety of methods, including tumor DNA and RNA sequencing and immunohistochemical staining. In patients with NTRK fusion positive tumors, NTRK inhibitors such as larotrectinib and entrectinib have shown good antitumor efficacy, with clinical response rates as high as 75%. Therefore, there is a need to improve the recognition and detection of fuch patients and to improve their prognosis by individualized and precise treatment with TRK inhibitors.

Expression of Wip1 in serum of patients with small cell lung cancer and its clinical significance / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of wild type p53 induced phosphatase 1 (Wip1) in small cell lung cancer (SCLC) cells and the serum of SCLC patient and its relationship with clinical prognosis. Methods: Real time quantitative PCR (qPCR) was used to detect the expression of Wip1 in SCLC cells and serum samples. Results: The expression of Wip1 in drug-resistant SCLC cells was significantly higher than that in sensitive cell lines (P<0.01). The expression of Wip1 in serum of SCLC group was significantly higher than that of normal control group (P<0.05); the expression of Wip1 in serum of patients with chemotherapy resistance was significantly higher than that in patients with chemotherapy sensitivity (all P<0.05); the serum Wip1 level was correlated with disease stage, chemotherapy sensitivity and survival status of SCLC patients (all P<0.05). The area under ROC curve of Wip1 predicting the prognosis of SCLC was 0.836 (95%CI:0.8230-0.9600, P<0.01); the expression lever of Wip1 was significantly correlated with progression free survival and overall survival time of SCLC patients (all P<0.05). Disease stage, chemosensitivity and Wip1 expression were independent prognostic factors for SCLC patients (all P<0.05). Conclusion: The expression of Wip1 in serum of SCLC patients may be related to chemotherapy sensitivity and prognosis. Wip1 may be a potential biomarker for therapeutic efficacy and prognosis evaluation of SCLC patients.

Genetic characterization of varicella zoster virus in Jilin province in 2017, China / 中华实验和临床病毒学杂志

Objective@#To clarify the genotype of varicella-zoster virus (VZV) in Jilin province in 2017, and to discriminate between vaccine strain and wild-type strain.@*Methods@#Vesicle fluid and throat swab samples were collected from 10 individuals with suspected VZV in Jilin province from January to March of 2017. Real-time fluorescent quantitative PCR was used to detect viral nucleic acid. Specific regions of ORF22, ORF38 and ORF62 of VZV were amplified by PCR. Viral genotype was determined by five SNPs of ORF 22 and vaccine strain or wild-type strain was distinguished by four SNPs of ORF 38 and ORF 62. The results were analyzed with MEGA5 and BioEdit software, using the VZV reference strain sequences from GenBank.@*Results@#VZV-positive strains were detected in 10 samples, all belonged to Clade 2. There was a synonymous mutation (C→T) in position 38 048 of JL17-7 strain. The nucleotide homology of ORF22 showed that all 10 samples were on the same branch with the Clade 2 referenced strains. Compared with Clade 2 referenced strains, the homology of nucleotide and amino acid for all 10 samples were 99.5%-100% and 99.3%-100%, respectively. The four specific SNPs of ORF38 and ORF62 in 10 samples were A-T-T-T, which were consistent with wild-type strain.@*Conclusions@#This study reveals that the VZV strains circulating in Jilin province in 2017 were all wild-type strains belonging to Clade 2.

Role of wild-type p53-induced phosphatase 1 in cancer / 国际外科学杂志

Wild-type p53-induced phosphatase 1 is a member of the protein phosphatase family and is an established oncogene due to its dephosphorylation of key protein in pathways and negative control of the DNA damage response system.Wild-type p53-induced phosphatase 1 serves a major role in tumorigenesis,progression,invasion,distant metastasis and chemotherapy resistance in various types of human cancer.Therefore,it may be a potential biomarker and therapeutic target in the diagnosis and treatment of cancer.In the paper,the current knowledge on the role of wild-type p53-induced phosphatase 1 in cancer is discussed.

1H-NMR based metabolomic analysis of chemical compositions in cultivated and wild pseudostellariae radix / 中国药学杂志

OBJECTIVE: To investigate the differences in the accumulation and biosynthesis of metabolites in different lactic acid of Pseudostellariae Radix. METHODS: 1H-NMR based metabolomic approach combined with multivariate statistical analysis was used to investigate the chemical compositions and find the different metabolites in cultivated Pseudostellariae Radix and its wild-type. RESULTS: Thirty-five metabolites were identified in the 1H-NMR spectra, and 15 of which showed remarkable differences. The multivariate statistical analysis showed that the cultivated Pseudostellariae Radix and its wild-type could be distinguished obviously. The contents of isoleucine, linolenic acid, lactic acid, alanine, lysine, glutamic acid, glutamine, acetoacetic acid, succinic acid, γ-aminobutyric acid, phenylalanine, and sucrose in cultivated Pseudostellaria heterophylla were lower than its wild-type, while the cultivated Pseudostellariae Radix contained more xylose, raffinose, and fumaric acid. CONCLUSION: This study will provide basic information for exploring the quality formation mechanism and revealing the accumulation and biosynthesis of metabolites in different ecotypes of Pseudostellariae Radix.

Possible mechanism and significance of rare p53 mutation in Epstein-Barr virus-associated gastric cancer / 中国肿瘤临床

In 2014, The Cancer Genome Atlas firstly classified gastric cancer into four types according to genotype. Epstein-Barr virus (EBV) positive gastric cancer or EBV-associated gastric cancer (EBVaGC) is attracting attention because it is a possibly suitable group for immunotherapy. Among the mutations observed in tumors, such as gastric cancer, p53 mutations are the most frequent. In particular, it occurs more frequently in EBVaGC than in EBV-negative gastric cancer (EBVnGC). Meanwhile, EBV infection is considered as an early event of tumorigenesis. The interactions between wild-type p53 proteins and BZLF1 (Z) proteins are essential in maintaining the latent state of EBV infection and promoting early replication. In the latter stages of replication, wild-type p53 proteins are degraded through the ubiquitination of some viral molecules. These findings may indicate the importance of wild-type p53 genes in EBVaGC formation. Inflammatory responses induced by EBV infection, tumor with a large number of lymphocyte infiltration, genome high mutation, and PD-L1 amplification make it possible to become the appropriate group of immunotherapy, which also illustrate that the important role of immune microenvironment during tumor progression. In EBVnGC, extremely high levels of p53 mutation were observed because of several associated factors, and the p53 protein encoded by the mutant p53 gene lost its antitumor function after tumorigenesis. In this review, the possible mechanisms of rare p53 mutation in EBVaGC are summarized.

iTRAQ-based quantitative proteomics of cultivated Pseudostellaria heterophylla and its wild type / 药学学报

This work was designed to investigate proteins differentially expressed in cultivated Pseudostellaria heterophylla and its wild type using iTRAQ proteomics approach. The extracted proteins were digested using FASP method and identified by iTRAQ coupled with LC-MS/MS technology and then analyzed by Protein Pilot 5.0 search engine. Proteins differentially expressed were searched through comparison of relatively quantified proteins. The analysis was conducted using GO (gene ontology), KEGG and STRING. A total of 3 775 proteins were detected, among them, 3 676 proteins can be quantified, of which 127 proteins were up-regulated and 205 were down-regulated in cultivated Pseudostellaria heterophylla. We found 71 significantly differentially expressed proteins for further analysis. These proteins were classified into nine categories:heat shock proteins, transferases, oxidoreductases, lyases, isomerases, ligases, hydrolases, tubulin and translocases. The results indicated that the carbohydrate and cellular amino acids metabolism of cultivated Pseudostellaria heterophylla were weaker than its wild type and its ability of responding to stress was much stronger. GWD1, PHS1, GBE1, PGM, and BAM1 are the important proteins to regulate sucrose; metE and CYS are the key proteins that regulate amino acids in cultivated and wild Pseudostellaria heterophylla. This will provide the basic information for exploring the cause of secondary metabolites differences in different ecotype of Pseudostellaria heterophylla and the protein mechanism of its quality formation.

Frecuencia y perfil de sensibilidad in vitro de aislamientos del Complejo Candida parapsilosis provenientes de pacientes con candidemias / Frequency and in vitro susceptibility profile of Candida parapsilosis Complex isolates from patients with candidemia

El objetivo de este trabajo fue conocer la frecuencia y el perfil de sensibilidad in vitro de aislamientos del Complejo Candida parapsilosis provenientes de casos de candidemias. Se estudiaron 754 cepas (Periodo 2008-2011), de la Red de Vigilancia de Candidemia del Instituto Nacional de Higiene “Rafael Rangel”. La identificación de las cepas se realizó por pruebas fenotípicas. La sensibilidad in vitro a los antifúngicos se evaluó por el método de Etest® y se determinó la concentración mínima inhibitoria a anfotericina B (AB), caspofungina (CS), fluconazol (FZ), y voriconazol (VZ). Se calcularon los puntos de corte epidemiológicos (PCE) y los rangos de cepas salvajes (PS) para cada antifúngico. El 43,6% de las cepas (n=328) fueron identificadas como Complejo C. parapsilosis; todas fueron sensibles a AB y presentaron bajos porcentajes de resistencia a FZ (4,3%), VZ (1,2%) y CS (0,6%). Los PCE y los rangos de PS (en µg/mL) fueron: FZ: 2/0,03-2; VZ y AB: 0,06/0,002-0,06 y CS: 0,5/0,002-0,5 respectivamente. Los resultados de este estudio aportaron información importante sobre el comportamiento del Complejo C. parapsilosis frente a los antifúngicos más utilizados en el tratamiento de las candidemias.

The aim of this study was to determine the frequency and in vitro susceptibility profile of Candida parapsilosis Complex isolates from patients with candidemia. Seven hundred and fifty four (754) strains (Period 2008-2011), from the Candidemia Surveillance Network of the Instituto Nacional de Higiene “Rafael Rangel” were studied. The strains identification was performed by phenotypic methods. In vitro antifungal susceptibility was evaluated by the Etest® method and minimum inhibitory concentration for amphotericin B (AB), caspofungin (CS), fluconazole (FZ), and voriconazole (VZ) was determined. Epidemiological cut off values (ECV) and ranges for wild type strains (WT) were also calculated for each antifungal. Forty three point six (43.6%) of the isolates (n=328) belonged to C. parapsilosis Complex; all of them were susceptible to AB and showed low resistance percentages to FZ (4.3%), VZ (1.2%) and CS (0.6%). The ECV and WT strains ranges (in mcg/mL) were: FZ: 2/0.03-2; VZ and AB: 0.06/0.002-0.06 and CS: 0.5/0.002-0.5 respectively. The results of this study provided important information about the behavior of the C. parapsilosis Complex against the most commonly antifungal agents used for the treatment of candidemias.

Distribución de la concentración inhibitoria mínima y puntos de corte "wild-type" en bacterias de origen animal en Argentina / Wild-typeminimal inhibitory concentration distributions in bacteria of animal origin in Argentina

El objetivo de este estudio fue determinar los patrones de resistencia antimicrobiana en bacterias indicadoras aisladas de muestras fecales de animales domésticos. La concentración inhibitoria mínima (CIM) fue determinada por el método de dilución en agar. El criterio de interpretación usado se basó en la distribución de la CIM y el punto de corte epidemiológico (ECOFF o ECV) de acuerdo con los datos del European Committee on Antimicrobial Susceptibility Testing (EUCAST). Los resultados obtenidos de 237 aislamientos de Escherichia coli mostraron sensibilidad reducida a ampicilina, estreptomicina y tetraciclina, antimicrobianos comúnmente usados en porcinos y aves de explotación intensiva. Con respecto a todas las especies del género Enterococcus spp., solo existe ECOFF o ECV para la vancomicina. De los 173 Enterococcus spp. aislados, sólo uno presentó sensibilidad reducida a dicho agente y fue categorizado como población 'non-wild-type' (NWT). Este es el primer informe en Argentina que presenta datos de puntos de corte epidemiológico en bacterias animales

The aim of this study was to determine the antimicrobial resistance profiles of indicator bacteria isolated from domestic animal feces. Minimal inhibitory concentration (MIC) was determined by agar dilution. Interpretative criteria on the basis of wild-type MIC distributions and epidemiological cutoff values (ECOFF or ECV) were used according to the 'European Committee on Antimicrobial Susceptibility Testing' (EUCAST) data. Results from 237 isolates of Escherichia coli showed reduced susceptibility for ampicillin, streptomycin and tetracycline, the antimicrobials commonly used in intensive breeding of pigs and hens. Regarding all the species of the genus Enterococcus spp., there are only ECOFF or ECV for vancomycin. Of the 173 Enterococcus spp. isolated, only one showed reduced susceptibility to vancomycin and was classifi ed as 'non-wild-type' (NWT) population. This is the fi rst report in Argentina showing data of epidemiological cutoff values in animal bacteria

The clinical efficacy of FOLFOX combined with bevacizumab/cetuximab in the first line treatment of advanced colorectal cancer with wild-type KRAS / 肿瘤

Objective: To compare the efficacy and main side effects of bevacizumab vs cetuximab plus FOLFOX regimen as the first-line treatment in advanced colorectal cancer patients with wild-type KRAS gene, and to evaluate the survival. Methods: Between January 2008 and February 2014, 72 patients were pathologically diagnosed of stage IV colorectal cancer with wild-type KRAS gene in People's Liberation Army General Hospital. Of 72 patients, 28 received FOLFOX regimen, 17 received bevacizumab combined with FOLFOX regimen, and 27 received cetuximab combined with FOLFOX regimen. After three cycles of chemotherapy, the short-term response was evaluated; the side effects were observed. All the patients were followed-up, and the progression-free survival (PFS) was calculated. Results: In FOLFOX regimen group, the objective response rate (ORR), disease control rate (DCR) and median PFS were 14.3%, 75.0% and 8.0 months, respectively; in bevacizumab combined with FOLFOX regimen group, these outcome measurements were 64.7%, 94.1% and 10.0 months, respectively; and in cetuximab combined with FOLFOX regimen group, these outcome measurements were 59.3%, 92.6% and 9.2 months, respectively. The ORR of FOLFOX regimen group was significantly lower than those in bevacizumab/cetuximab combined with FOLFOX regimen groups (χ2 = 12.101, P= 0.000 5; χ2 = 12.014, P = 0.000 5). There were no significant differences in DCR and median PFS among the three groups (P > 0.05). Conclusion: Bevacizumab/cetuximab combined with FOLFOX regimen can improve the ORR, DCR and median PFS in advanced colorectal cancer patients with wild-type KRAS. These two combined regimens have a quite similar efficacy, less adverse reactions and good tolerance.

Constructing a p53-fused dual luciferase reporter and verifying its function / 中南大学学报(医学版)

Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter. Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively. Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.

Antibody-directed lentiviral vector-mediated hepatocarcinoma cells targeting IGF1R- siRNA/P53 genes therapy / 中华肝胆外科杂志

ObjectiveTo investigate the targeting infection of single chain antibody againstAFP (scFv anti-AFP) directed lentivirus and the inhibitory effects of a dual-growth inhibition systemon hepatocarcinoma cells.MethodsPlasmids WtP53-pPRIME-miR30-shRNA-IGF1R,pMD2G-Anti-AFP,and psPAX2 have previously been constructed to cotransfect to the packaging cell line 293Tusing Lipofectamine2000.The infection results were observed through fluorescence microscopy.PCRand Western blotting were used to demonstrate the successful transduction and transcription of theWtP53-pPRIME-miR30-shRNA-IGF1R gene.The effects of reconstructed lentivirus infected liver cellgrowth were assessed by the cell growth curve of CCK8 cells. Apoptosis was evaluated by theTUNEL assay.ResultsRecombined lentivirus was successfully constructed with the functional PFUtiters of recombined lentivirus at 4.58× 109PFU/ml.This positive result was confirmed by PCR andWestern blotting.ConclusionsThe targeted therapy mediated by anti-AFP scFv could significantlyinhibit the proliferation of HEP3B cells and promote the apoptosis.

Inhibition of subretinal co-transfection of Rb94 and wild-type p53 gene on retinoblastoma by ultrasound microbubble in nude mouse / 中华实验眼科杂志

Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80％ (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P＜0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.

Rotavirus A genotype G1P[8]: a novel method to distinguish wild-type strains from the Rotarix® vaccine strain

Rotaviruses are important enteric pathogens for humans and animals. Group A rotaviruses (RV-A) are the most common agents of severe gastroenteritis in infants and young children and vaccination is the most effective method to reduce RV-A-associated diseases. G1P[8], the most prevalent RV-A genotype worldwide, is included in the RV-A vaccine Rotarix®. The discrimination between wild-type G1P[8] and vaccine G1P[8] strains is an important topic in the study of RV-A epidemiology to manage outbreaks and to define control measures for vaccinated children. In this study, we developed a novel method to segregate the wild-type and vaccine strains using restriction endonucleases. The dsRNA from the Rotarix® vaccine was sequenced and the NSP3 gene was selected as the target gene. The vaccine strain has a restriction pattern that is different than that of wild-type RV-A G1P[8] isolates after digestion with the restriction endonuclease BspHI. This pattern could be used as a marker for the differentiation of wild-type G1P[8] strains from the vaccine strain.

EST sequencing and SSR marker development from cultivated peanut (Arachis hypogaea L)

Making use of the gene resources of wild type peanuts is a way to increase the genetic diversity of the cultivars. Marker assisted selection (MAS) could shorten the process of inter-specific hybridization and provide a possible way to remove the undesirable traits. However, the limited number of molecular markers available in peanut retarded its MAS process. We started a peanut ESTs (Expressed Sequence Tags) project aiming at cloning genes with agronomic importance and developing molecular markers. In this study we found 610 ESTs that contained one or more SSRs from 12,000 peanut ESTs. The most abundant SSRs in peanut are trinucleotides (66.3 percent) SSRs and followed by dinucleotide (28.8 percent) SSRs. AG/TC (10.7 percent) repeat was the most abundant and followed by CT/GA (9.0 percent), CTT/GAA (7.4 percent), and AAG/TTC (7.3 percent) repeats. Ninety-four SSR containing ESTs were randomly selected for primer design and synthesis, of which 33 pairs could generate good amplification and were used for polymorphism assessment. Results showed that polymorphism was very low in cultivars, while high level of polymorphism was revealed in wild type peanuts.

Antitumor metabolites produced by two Penicillium purpurogenum G59 mutants / 国际药学研究杂志

Objective: To investigate the antitumor metabolites of fungal mutants 2-2-3 and PDN-f-2, the two bioactive mutants of Penicillium purpurogenum G59 that do not produce antitumor metabolites. Methods: Bioactive metabolites newly produced by the mutants were isolated by a bioassay-guided separation procedure using iquid-liquid extraction, column chromatography and recrystallization methods through direct comparison with the sample from P. purpurogenum G59. The compounds obtained were identified by spectroscopic methods. The antitumor activity was assayed by the MTT method using K562 cells. Results: Two bioactive metabolites 1 and 2 were isolated from the fermentation products of 2-2-3 and PDN-f-2, respectively, and identified as ergone (1) and citrinin (2). Compounds 1 and 2 inhibited the proliferation of K562 cells with the IC50 values of 7. 4 and 48. 0 μg/ml, respectively. Conclusion: Compounds 1 and 2 are the antitumor metabolites newly produced by he mutants 2-2-3 and PDN-f-2, respectively, and have not been found in the metabolites of P. purpurogenum so far. It is revealed from the present result that the alteration of secondary metabolism of wild-type fungal strains without bioactivity for obtaining bioactive metabolite-producing mutants may become a new route to expand the source of new fungal strains for drug screening.

Effect of Wild Type p53 Gene on Inhibiting the Centrosome Hyperamplification in Bladder Cancer Cell Line / 中国医科大学学报

Objective To study the effect of wild type p53 gene on centrosome hyperamplification in bladder cancer cells.Methods A wild type p53 gene recombinant adenovirus vector AdCMVp53 was constructed,and then trangfected into the human bladder cancer cell line T24.The cells were stained with the monoclonal antibody against pericentrin by indirect immunofluorescence method.The change of centrosome hyperamplification was observed under the fluorescence microspcope.Results Introduction of wild type p53 could suppress the centrosome amplification of T24 cell line.Conclusion p53 might play an important role in the regulation of centrosome hyperamplification.The loss of p53 might be one of the mechanisms involved in chromosome instability and contribute to the genesis and development of the bladder carcinoma.

Expression and Significance of Cyclin A1 mRNA in the Skin of Wild-Type Mice / 中国实验动物学报

Objective To study the expression and significance of cyclin A1 in the skin of wild-type mice at RNA level.Methods Thirty 6-12-week old wild-type Kunming mice(15 male and 15 female)were included in this study.In situ hybridization was used to detect the expression of cyclin A1 mRNA in the skin of head and neck of the mice.The skin treated without probe was regarded as negative control and the testis of male mice was taken as positive contr01.Results The expression of cyclin A1 mRNA was found in sebaceous glands of 25 mice and in epidermis of 12 mice.Strong positive staining of sebaceous glands was seen in 50% and positive staining in 33.3% of sections,whereas strong positive and positive staining of epidermis was seen in 13.3% and 20% of sections.respectively.The positive rate of sebaceous glands was 83.3%,much higher than that of epidermis(33.3%).Conclusions There is a quite high expression of eyelin A1 mRNA in the skin sebaceous gland and epidermis of head and neck of wild-type mice.Especially a strong expression is in the sebaceous glands.It indicates that cyclin A1 mRNA may play a certain role in the physiological function of sebaceous glands and epidermis of the skin.