Acinetobacter Infections ; microbiology ; Acinetobacter baumannii ; drug effects ; isolation & purification ; Anthracosis ; microbiology ; Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; Fosfomycin ; pharmacology ; Humans ; Imipenem ; pharmacology ; Pneumonia, Bacterial ; microbiology

In order to have a better understanding of the species diversity of medical plants in Enshi, Hubei of China, extensive field investigations and specimen collections were conducted in Enshi and adjacent regions. Based on field observations of plants in their living habitats and comparative morphological studies on specimens in herbarium of Hubei minzu University and other available herbaria as well, three new records of medical plants in Hubei, Scutellaria yunnanensis, Alangium faberi var. heterophyllum, and Drymaria diandra, were reported in this paper.
China ; Plants, Medicinal ; Records as Topic

OBJECTIVETo elucidate the relationship between point mutations of nt3243A --> G, nt3426 A --> G of mitochondrial DNA and type 2 diabetes mellitus(DM).

METHODSTwo hundred patients with type 2 DM and 180 controls with normal glucose tolerance and absence of DM family history were included. The mutations were determined by PCR-restriction fragment length polymorphism.

RESULTSThe point mutation nt3426A --> G of mitochondrial DNA ND1 was found in 2 of the patients with type 2 DM (1.0%) but in none of the controls (0). The incidence of this mutation showed no significant difference between the two groups(P>0.05). And none was found to have the mutation of nt3243 --> G.

CONCLUSIONThe point mutation nt3426 A --> G of mitochondrial DNA ND1 may not be an independent factor to cause type 2 DM.

Adult ; Aged ; Aged, 80 and over ; DNA, Mitochondrial ; analysis ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; Family ; Humans ; Male ; Middle Aged ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Young Adult

BACKGROUNDRecent studies have indicated that many mutations in mitochondrial (mt) DNA NDI gene region are related to diabetes mellitus. In this study we explored the relationship between various mtDNA ND1 gene mutations and type 2 diabetes mellitus (DM) among Chinese.

METHODSUsing PCR restriction fragment length polymorphism (PCR-RFLP) analysis and gene sequencing, 4 spots of mtDNA (nt3243, nt3316, nt3394, nt3426) were screened in 478 diabetics and 430 non-diabetic subjects.

RESULTSIn diabetic group, there were 13 carriers (2.72%) of 3316 G-->A mutation,12 (2.51%) of 3394 T-->C mutation and 2 (0.42%) of 3426A-->G mutation. In controls, only 3394 T-->C mutation was observed in 2 subjects (0.47%). There was significant difference in the frequency of 3316 and 3394 mutation between two groups (P < 0.05, respectively). More subjects with mitochondrial DNA ND1 gene mutations had DM family history and greater tendency of maternal inheritance when compared to those patients without mutation in diabetic group (P < 0.01). A 3426 mutation diabetic pedigree was studied, and we found 12 maternal members in the family had the same mutation.

CONCLUSIONmtDNA ND1 gene mutations at nt3316 (G-->A), nt3394 (T-->C) and 3426 (A-->G) might contribute to the pathogenesis of DM with other genetic factors and environment factors.

Base Sequence ; DNA, Mitochondrial ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; Humans ; Molecular Sequence Data ; Mutation ; NADH Dehydrogenase ; genetics ; Pedigree ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA

Objective To evaluate the effect of health education in controlling the iodine deficiency diserders(IDD) in order to provide reference data for the further prevention and control. Methods Each village of 3 towns in Congjiang County was selected in 2007, where the health education lasting for 10 months had been implemented in the school students of 3-6 grade and the villagers. The school students of 3-6 grade and 30 housewives in the villagers were investigated for their IDD control knowledge, the salt consuming conditions as well as the sales of both rough and fine salt at a salt retail site in each village before and after the health education was implemented. Results The awareness rate of the knowledge of IDD control in the students and housewives was 91.4% (581/636) and 78.3% (282/360), respectively after intervention, which significantly increased (χ2= 532.044, 326.117, both P < 0.01) compared with the rate of 28.2% (184/652) and 11.4% (41/360) before intervention. The proportion of consuming fine salt was 91.8%(146/159) and 95.6%(86/90), significantly inereased(χ2= 236.623, 135.350, both P < 0.01) compared with 6.1%(10/163) and 7.8% (7/90) found before intervention. The selling proportion of fine salt at the salt retail site in the village was 60.0%(900/1500), significantly increased(χ2= 824.176, P < 0.01) compared with 10.0%(150/1500) before intervention. Conclusions Health education and promotion is solid foundation for effectively controlling IDD, through which the students and villagers are actively and voluntarily involved in the program and hence have formed good living and hygiene habits, thus expected effect has been obtained.

Adult ; China ; Genes, Viral ; Genotype ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; Methadone ; therapeutic use ; Middle Aged ; Opioid-Related Disorders ; virology

OBJECTIVETo obtain human mitochondrial transcription factor A (TFAM), mitochondrial transcription factor B1 (TFB1M), and mitochondrial transcription factor B2 (TFB2M) that were expressed efficiently in E. coli BE21 and to purify the target proteins.

METHODSTFAM, TFB1M, and TFB2M segments were designed and synthesized. After having been sequenced, the reconstructed expression vectors were constructed by enzyme digestion and by cloning into an expression vector pET42a. Then the reconstructed vectors were transformed into E. coli BL21. Recombinant glutathione S transferase (GST) fusion proteins were expressed via the induction of IsoPropyl beta-D-ThioGalactoside (IPTG) and purified by glutathione Sepharose 4B.

RESULTSThe expression plasmids of pET42a-TFAM, pET42a-TFB1M, and pET42a-TFB1M were successfully constructed. The sequences of the cloned gene segments were identical with GenBank reported. The protein bands with relative molecular masses of 56 000, 67 000, and 69 000 appeared on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) after the expressed GST-TFAM, GST-TFB1M, and GST-TFB2M fusion proteins were separated by SDS-PAGE. The expressed fusion proteins were purified to high purity.

CONCLUSIONThe recombinant plasmids pET42a-TFAM, pET42a-TFB1M, and pET42a-TFB2M were successfully constructed, and the GST-fused target proteins were prepared.

Cloning, Molecular ; DNA-Binding Proteins ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Methyltransferases ; genetics ; Mitochondrial Proteins ; genetics ; Recombinant Fusion Proteins ; genetics ; Transcription Factors ; genetics

The silent information regulator protein 2 (Sir2) and its homologues play an important role in the regulation of cellular physiological processes such as survival, apoptosis, and aging. SIRT1, the mammalian Sir 2 homologue, has been shown to deacetylate a wide range of non-histone substrates and histone substrates. It has been constantly reported that SIRT1 may be associated with the occurrence of metabolic syndrome, genomic homeostasis, tumors, and neurodegenerative diseases. Calorie restriction may mitigate many major diseases in rodent models by SIRT1-mediated deacetylase activity and prolong the life expectancies in these animals. Therefore, SIRT1 may be emphasized as a new therapy target for many different diseases.
Animals ; Caloric Restriction ; Humans ; Longevity ; Sirtuin 1 ; genetics ; physiology ; Substrate Specificity

OBJECTIVETo study the regulatory mechanism of SATB1 repression in cells other than T cells or erythroid cells, which have high expression level of SATB1.

METHODSHeLa epithelial cells were treated with either histone deacetylase inhibitor (HDACi) trichostatin A (TSA) or DNA methylation inhibitor 5-Aza-C before detecting SATB1 expression. Luciferase reporter system was applied to measure effects of EZH2 on SATB1 promoter activity. Over-expression or knockdown of EZH2 and subsequent quantitative reverse transcription-polymerase chain reaction were performed to determine the effect of this Polycomb group protein on SATB1 transcription. Chromatin immunoprecipitation (ChIP) assay was applied to measure enrichment of EZH2 and trimethylated H3K27 (H3K27me3) at SATB1 promoter in HeLa cells. K562 cells and Jurkat cells, both having high-level expression of SATB1, were used in the ChIP experiment as controls.

RESULTSBoth TSA and 5-Aza-C increased SATB1 expression in HeLa cells. Over-expression of EZH2 reduced promoter activity as well as the mRNA level of SATB1, while knockdown of EZH2 apparently enhanced SATB1 expression in HeLa cells but not in K562 cells and Jurkat cells. ChIP assay Results suggested that epigenetic silencing of SATB1 by EZH2 in HeLa cells was mediated by trimethylation modification of H3K27. In contrast, enrichment of EZH2 and H3K27me3 was not detected within proximal promoter region of SATB1 in either K562 or Jurkat cells.

CONCLUSIONSATB1 is a bona fide EZH2 target gene in HeLa cells and the repression of SATB1 by EZH2 may be mediated by trimethylation modification on H3K27.

Azacitidine ; pharmacology ; Base Sequence ; Cell Line ; Chromatin Immunoprecipitation ; DNA Methylation ; DNA Primers ; DNA-Binding Proteins ; physiology ; Enhancer of Zeste Homolog 2 Protein ; Epigenesis, Genetic ; physiology ; Epithelium ; metabolism ; Gene Silencing ; Humans ; Hydroxamic Acids ; pharmacology ; Matrix Attachment Region Binding Proteins ; genetics ; Polycomb Repressive Complex 2 ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; physiology

OBJECTIVETo investigate whether α-hemoglobin stabilizing protein (AHSP), the α-globin-specific molecular chaperone, is regulated by erythroid transcription factor NF-E2.

METHODSWe established the stable cell line with NF-E2p45 (the larger subunit of NF-E2) short hairpin RNA to silence its expression. Western blot, real-time polymerase chain reaction, and chromatin immunoprecipitation (ChIP) analysis were performed to detect the expression of AHSP, the histone modifications at AHSP gene locus, and the binding of GATA-1 at the AHSP promoter with NF-E2p45 deficiency. ChIP was also carried out in dimethyl sulfoxide (DMSO)-induced DS19 cells and estrogen-induced G1E-ER4 cells to examine NF-E2 binding to the AHSP gene locus and its changes during cell erythroid differentiation. Finally, luciferase assay was applied in HeLa cells transfected with AHSP promoter fragments to examine AHSP promoter activity in the presence of exogenous NF-E2p45.

RESULTSWe found that AHSP expression was highly dependent on NF-E2p45. NF-E2 bound to the regions across AHSP gene locus in vivo, and the transcription of AHSP was transactivated by exogenous NF-E2p45. In addition, we observed the decrease of H3K4 trimethylation and GATA-1 occupancy at the AHSP gene locus in NF-E2p45-deficient cells. Restoration of GATA-1 in G1E-ER4 cells in turn led to increased DNA binding of NF-E2p45.

CONCLUSIONNF-E2 may play an important role in AHSP gene regulation, providing new insights into the molecular mechanisms underlying the erythroid-specific expression of AHSP as well as new possibilities for β-thalassemia treatment.

Base Sequence ; Blood Proteins ; genetics ; DNA Primers ; GATA1 Transcription Factor ; physiology ; Gene Expression Regulation ; physiology ; Gene Silencing ; HeLa Cells ; Humans ; Methylation ; Molecular Chaperones ; genetics ; NF-E2 Transcription Factor, p45 Subunit ; physiology ; Promoter Regions, Genetic ; Reverse Transcriptase Polymerase Chain Reaction