To study the chemical components and the antifungal activity of extraction from conidia of Trichoderma viride LTR-2.The extraction were obtained by distilling with Methylene dichloride from conidia of Trichoderma viride LTR-2 cultured on wheat bran solid matrix.Antifungal activity were determined by mycelium growth method.The chemical components of the extraction were analysed by GC-MS,the relative components in the extraction were determined by area normalization.The extraction not only have broad-spectrum control,showed antibiosis against eleven different plant fungal pathogens in PDA dish,such as Rhizoctonia solani,Alternaria brassica,Verticillium dahliae,Macrophoma kawatsukai,Fusarium moniliforme,Botrytis cinerea,Rhizoctonia cerealis,Fusarium oxysporum f.sp.vasinfectum,Bipolaris sorokinana,Fusarium graminearum,Alternaria.mali,but also have high inhibitory effect,and had 89.3% suppressive rate to Rhizoctonia cerealis.About sixty components were separated and identified by GC-MS,majority components were Hydrocarbon,the number of the Hydrocarbon were fourty-three kinds.Ergosterol was the major chemical components of the extract,and has 41.90% content.Other components comprised:Ketone,Organic acid,Alcohol,Ene,et al.Conclusion:The extraction from conidia of Trichoderma viride LTR-2 have antifungal activity.The extration comprised 2H-Pyran-2-one,5,6-dihydro-6-pentyl,it has 2.35% content.reference others literature,2H-Pyran-2-one,5,6-dihydro-6-pentyl may be the suppressive component of the extration.

Autopsy ; Female ; Hepatoblastoma ; diagnosis ; diagnostic imaging ; pathology ; Humans ; Infant, Newborn ; Liver Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Neoplasm Staging ; Rare Diseases ; Tomography, X-Ray Computed

AIMTo study the protective effect of sodium pyruvate on ischemia/reperfusion injury following hemorrhagic shock.

METHODSRat models of hemorrhagic shock were built up. When the shed blood was infused, the rats were also randomly provided by one of normal saline, glutathione and sodium pyruvate. Rats were killed 3 hours after the reperfusion, the activity of plasma lactate dehydrogenase (LDH) and glutamic-oxaloacetic transaminase (GOT), the level of tissue malondialdehyde (MDA) and the activity of tissue myeloperoxidase (MPO) were detected. Biopsy specimens were obtained to investigate morphological changes of the myocardial, hepatic, lung and renal tissue.

RESULTSThe activity of plasma LDH and GOT, the level of MDA of hepatic, lung and renal tissue and the activity of MPO of myocardial, lung and renal tissue decreased remarkably in group given sodium pyruvate compared with group given normal saline, and the effect of group given sodium pyruvate was more remarkable than group given glutathione.

CONCLUSIONThese data support the view that sodium pyruvate shows protective effect on ischemia/reperfusion injury following hemorrhagic shock. It is possibly relevant to scavenging of oxygen free radicals, reduction of neutrophil, and anti-inflammatory response.

Animals ; Aspartate Aminotransferases ; blood ; Disease Models, Animal ; Kidney ; metabolism ; pathology ; L-Lactate Dehydrogenase ; blood ; Liver ; metabolism ; pathology ; Lung ; metabolism ; pathology ; Male ; Malondialdehyde ; analysis ; Peroxidase ; analysis ; Protective Agents ; pharmacology ; Pyruvic Acid ; pharmacology ; Rats ; Rats, Wistar ; Reperfusion Injury ; blood ; pathology ; prevention & control ; Shock, Hemorrhagic ; blood ; pathology

Streptomyces hygroscopicus 17997 produces the antiviral and antitumor ansamycin antibiotic, geldanamycin. Studies on geldanamycin biosynthetic pathway will provide good tools for genetic manipulation of the antibiotic-producing strain to improve the productivity or to facilitate making novel geldanamycin analogs. The structural similarities between geldanamycin and ansamycins such as rifamycin or ansatrienin suggest that both geldanamycin and ansamycins has a closely related pathways of biosynthesis and that biosynthetic system for geldanamycin is similar to the one of type I polyketide synthase (PKS) enzyme system. To explore the possible PKS genes involved in geldanamycin biosynthesis, the degenerate primers were designed according to the conserved sequence of KS-AT region from erythromycin and oleandomycin type I PKS genes. Cosmids containing multiple PKS genes (pCGBK2,4,6,10,11,18) were obtained by hybridization with the PCR products, which were amplified from S. hygroscopisus 17997 genomic DNA. The designed primers above were used for PCR. Development of a Streptomyces temperate phage phiC31-derivative KC515( tsrR) transduction system was carried out for identification of cosmids containing the PKS gene related to biosynthesis of geldanamycin. Several factors, mainly the Ca2+ and Mg + concentrations in different culture media affecting the frequency of gene transfection, were optimized .Transfection efficiency could reach up to 10(3) /microg DNA on YMG medium supplemented with 10mmol/L MgSO4. Reversely, the transfection efficiency decreased when YMG medium was supplemented with 30mmol/L MgSO4. Gene transfection system based on the integration-defective phage KC515 had been established for S. hygroscopicus17997. Recombinant phages (ph111, 258, 287, 116, 105) were constructed by insertion of the homologous to PKS gene fragments into the KC515 phage vector. Gene disruption experiments were performed by transduction of recombinant phages into S. hygroscopicus 17997 genome, and disruption of geldanamycin production was observed as a result of homologous recombination between the cloned insert in recombinant phage and the S. hygroscopicus 17997 genome by integration. Thiostrepton resistant transductants were selected and integration event was analyzed by Southern hybridization. The fermentation broth extracts from five resistant transductants were analyzed by TLC and HPLC. The results showed that only G16 mutant failed to produce geldanamycin. This result showed that the integration of the insert DNA fragment in recombinant phage phl6 into the chromosome of S. hygroscopicus disrupted the expression of the geldanamycin biosynthetic genes. The original cosmid pCGBK10 containing this cloned insert was predicted to encode PKS genes in the geldanamycin biosynthesis. This study laid the foundation for cloning the PKS genes involved in geldanamycin biosynthetic gene cluster from S. hygroscopicus 17997.
Alcohol Oxidoreductases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Bacteriophages ; genetics ; Benzoquinones ; metabolism ; Blotting, Southern ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Genetic Vectors ; genetics ; Lactams, Macrocyclic ; metabolism ; Multigene Family ; genetics ; physiology ; Polymerase Chain Reaction ; Streptomyces ; genetics ; metabolism

Objective To study the inducement of U251 glioblastoma cell apoptosis in vivo through up-regulating PUMA expresion and knocking down miR-221/222, and explore its mechanism.Methods Nude mouse xenograft models were established in 5-week-old BALB/c nude mice by subcutaneous vaccination of U251 glioblastomas; 1 week later, they were treated with intratumoral injection of lipofcctamine-mediated miRNA-221/222 antisense oligonucleotides (GroupA), nonsense sequences (Group B) and controls (Group C),respectively (n=8).The tumor growth was monitored until the end of observation period (28 d after the treatment) and pathological changes of the glioblastoma tissues were observed by HE staining at the end of observation.Fluorescence in situ hybridization (FISH) and real-time PCR were employed to measure the miR-221 and miR-222 expressions. Terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling (TUNEL) assay was used to detect the apoptosis of glioblastomas.Immunohistochemistry and Westem blotting were used to detect the expressions of PUMA,bax,bcl-2 and p53 in removed tumor specimens. Results The volume in Group A was significantly smaller than that of those in group B and group C 6-28 dater treatment (P=0.006). The miR-221 and miR-222 mRNA expressions in Group A were significantly decreased as compared with those of those in group B and group C.HE staining indicated that decreased heteromorphism and reduced number of new vessels in Group A were noted as compared with those in group B and group C.The cell apoptotic index in Group A was significantly higher than that in group B and group C (P＜0.05).Immunohistochemistry showed that the expression levels of PUMA and bax in Group A was significantly up-regulated as compared with those in group B and group C, while the expression of bcl-2 in Group A was significantly down-regulated as compared with that in group B and group C; and no significant changes were noted in the p53 expression. Conclusion By up-regulating PUMA expresion,knocking down miR-221/222 can induce U251 glioma apoptosis in vivo.

Objective To investigate plasma glutathione S-transferase(GSTs) activity and GSTP1 gene Ile105Val polymorphism in Bijie City, Guizhou Province, a coal-burning fluorosis endemic area. Methods One hundred and sixty villagers from Yachi Twon using non-improved cooking stoves were selected as the non-intervened group in Bijie City, Guizhou Province where coal-burning fluorosis was prevailing; 153 villagers as the intervented group were chosen from Changchun Twon, where cooking stoves were improved; 151 villagers were served as the control group from Baiyunshan Twon, Changshun County without endemic fluorosis. The activity of GSTs was tested by colorimetric analysis with spectrophotometer. The genotype of the GSTP1 gene Ile105Val polymorphism, presenting as either homozygous wild-type (AA), or heterozygous mutation type (AG), or homozygous mutation type (GG), was detected through the PCR-RFLP procedure. Results The activity of GSTs in plasma of non-intervened group [(12.44±4.97) kU/L]was significantly lower than that of intervened group (P < 0.05), and that of intervened group[(20.78±6.20)kU/L]was significantly lower than that of control group[(24.30±6.27)kU/L, P< 0.05]. The difference of the enzyme activity of three groups were statistically significant (F = 51.71, P < 0.05), but this enzyme activity did not vary significantly in each sex of each grnup(P > 0.05). Compared intervened group [AA:67.3%(103/153), AG:29.4%(45/153),GG:3.3%(5/153)]and non-intervened group[AA:66.9%(107/160), AG:30%(48/160), GG:3.1%(5/160)]with control group[AA:74.8%(113/151), AG:25.2%(38/151), GG:0 (0/151)], the Ile105Val polymorphism site of GSTP1 gene had significant difference(χ2= 6.04,6.07, both P< 0.05), but not significant between intervened and non-intervened groups(χ2 = 0.02, P>0.05). Conclusions Fluorosis can decrease the activity of GSTs and introduce the GSTP1 gene Ile105Val polymorphism, intervention with the fluorine intake will improve the effect of fluoride on the body.

Background Qiang minority is minority groups of China with the special habits and customs and living condition. So whether the spectrum of disease and bacteria spectrum in conjunctiva are similar with Han nationality is worth paying attention. Objective Present survey was to obtain the data about bacterial species in conjunctival sac in Qiang minority population with the age 40 years old and more and the compare with matched Han nationality population. Methods This survey study was performed as the standardized training and protocol. A total of 212 eyes of 106 individuals from Qiang minority in Beichuan county and 640 eyes of 320 subjects from Han nationality in Mianyang city received questionnaire survey and ophthalmological examination. The secretion of the inferior palpebral conjunctival sac was embrocated and inoculated on blood plate for 48-72 hours. The bacteria was separated and identified. This study was approved by the Ethic Committee of Sichuan Provicial People' s Hospital. Orally informed consent was obtained before the medical procedure. Results All the examinee finished the survey and examination with a good compliance. No significant difference was found in the demography between these two groups of population. The multiple bacterial positive rate in conjunctival sac was 59. 4% in Qiang minority and that of Han people was 66. 3% with a considerably difference between them (χ2 = 2. 27,P = 0. 13). The multiple bacterial species were simultaneously detected in 26.2% in Qiang minority population and 11.88% Han people, showing evidently difference (χ2 = 106. 40, P = 0. 00 ) . The positive rate of corynbaccterium in conjunctival sac of Qiang minority was statistically lower than that of Han people (20. 7% versus 45. 0% ,χ2 =31. 75 ,P = 0. 00) ,but there was no statistical difference in the positive rate of staphylococcus epidemics between two groups (χ2 = 1. 89 ,P = 0. 17). Conclusion The bacteria positive rate in conjunctiva sac is resemble in the population over 40 years in both the Qiang minority and Han nationality. The simple bacterial species is found in majority people in two groups of subjects. The positive rate of multiple bacterial strains coexistence is more in the Qiang minority. The bacterial strains is different between Qiang minority and Han nationality.

Animals ; Brain ; drug effects ; metabolism ; Carbon Dioxide ; metabolism ; Chronic Disease ; Curcumin ; pharmacology ; Disease Models, Animal ; Fas Ligand Protein ; metabolism ; Hypoxia ; metabolism ; Male ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; metabolism

Objective To explore the relationship between social participation and needs for rehabilitation of the disabled in Guangdong Province and to make a proposal for developing the rehabilitation strategies. Meth-ods The data of the Second National Sample Survey of Disabled Persons in Guangdong Province was used in this study. Ranked data analysis was made with the sub-items of the social participation assessment and the main needs of the disabled individuals. Results Significantly statistical differences were revealed with regard to the constitu-ent ratio of needs for rehabilitation services among people with different degrees of difficulties in social participation caused by hearing and visual impairments as well as physical and mental disabilities. No significant difference was found in terms of the constituent ratio of rehabilitation needs among those with difficulties in speech and those with psychiatric diseases. The major rehabilitation needs focused on medical service, assistive apparatus support and functional trainings. Conclusions The rehabilitation needs were different among different categories of disabled persons. Rehabilitation services should be provided accordingly.