Identification and validation of a new target is one of the most important steps for new antituberculosis (TB) drug discovery. Researches have shown that Mycobacterium tuberculosis (Mtb) encodes 20 CYP450 enzymes which play important roles in the synthesis and metabolism of lipid, cholesterol utilization, and the electron transport of respiratory chain in Mtb. With the critical roles within the organism as well as the protein structures of six Mtb CYP450 enzymes being clarified, some of them have been highlighted as potential anti-tuberculosis targets. In this paper, the phylogenetic analysis, the structural features, and the enzymatic functions of Mtb CYPs, as well as the mechanism of interactions with selective inhibitors such as azole antifungal agents for the CYPs have been reviewed and summarized. The druggability of the CYPs has also been analyzed for their further utility as targets in high throughput screening and rational design of more selective inhibitors.
Antitubercular Agents ; chemistry ; pharmacology ; Azoles ; chemistry ; pharmacology ; Cytochrome P-450 Enzyme Inhibitors ; chemistry ; pharmacology ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Drug Delivery Systems ; methods ; Drug Discovery ; Humans ; Mycobacterium tuberculosis ; drug effects ; enzymology ; genetics ; Phylogeny ; Tuberculosis ; drug therapy ; microbiology

The ability of repair and regeneration of central nervous system (CNS) is limited. So many researchers applied themselves to search a valuable cell resource for treating severe diseases of the CNS. Several studies from different laboratories have recently reported that stem cells derived from human umbilical cord blood under certain in vitro conditions can manifest neural features that resemble features of neural-derived cells. In vivo transplantation studies have shown that these stem cells persistently engraft in the CNS, some engrafted cells acquire the characteristics of neurons and glia, and improve functional recovery after central nervous system injury. The existence of stem/progenitor cells with previously unappreciated proliferation and differentiation potential in umbilical cord blood raise the possibility that cord blood may provide an efficient source of cells differentiating into the neural lineage, with a potential to be employed in the therapy of human CNS diseases. The achievement and focuses on the mechanisms and modulation of induction of differentiation and in vitro and in vivo studies in this field was reviewed.

Identification and validation of a new target is one of the most important steps for new antituberculosis (TB) drug discovery. Researches have shown that Mycobacterium tuberculosis (Mtb) encodes 20 CYP450 enzymes which play important roles in the synthesis and metabolism of lipid, cholesterol utilization, and the electron transport of respiratory chain in Mtb. With the critical roles within the organism as well as the protein structures of six Mtb CYP450 enzymes being clarified, some of them have been highlighted as potential anti-tuberculosis targets. In this paper, the phylogenetic analysis, the structural features, and the enzymatic functions of Mtb CYPs, as well as the mechanism of interactions with selective inhibitors such as azole antifungal agents for the CYPs have been reviewed and summarized. The druggability of the CYPs has also been analyzed for their further utility as targets in high throughput screening and rational design of more selective inhibitors.

Objective To obtain the gene fragment encoding human mannan-binding lectin (MBL)-associated serine protease-2 (MASP-2). Methods The target cDNA fragment was amplified from the total RNA extracted from human fetal liver tissue by RT-nested PCR, cloned into pGEM-T vector and identified by restriction analysis and sequencing. Results The cDNA fragment encoding MASP-2 polypeptide with the signal sequence was isolated, linked with pGEM-T vector and transformed into E.coli TG1. The restriction maps of the selected clones were consistent with those generated by computer analyses. The result of sequencing of one selected clone showed that its sequence was identical with that of reported human MASP-2 cDNA. Conclusion The human MASP-2 gene was successfully cloned, which prepares the ground for studying the molecular mecha- nisms by which MBL activates the complement lectin pathway and the mutations of MBL gene cause immunodeficiency.

Objective To obtain the gene fragment encoding human mannan-binding lectin (MBL)-associated serine protease-2 (MASP-2). Methods The target cDNA fragment was amplified from the total RNA extracted from human fetal liver tissue by RT-nested PCR, cloned into pGEM-T vector and identified by restriction analysis and sequencing. Results The cDNA fragment encoding MASP-2 polypeptide with the signal sequence was isolated, linked with pGEM-T vector and transformed into E.coli TG1. The restriction maps of the selected clones were consistent with those generated by computer analyses. The result of sequencing of one selected clone showed that its sequence was identical with that of reported human MASP-2 cDNA. Conclusion The human MASP-2 gene was successfully cloned, which prepares the ground for studying the molecular mecha- nisms by which MBL activates the complement lectin pathway and the mutations of MBL gene cause immunodeficiency.

This study was aimed to evaluate the effect of triptolide (TPL) on the reversal of multidrug resistance in K562/A02 cell line. The sensitivity of K562 and K562/A02 to adriamycin (ADM) and reversal of drug resistance were determined with MTT method. The concentration of intracellular ADM and P-glycoprotein expression were detected by flow cytometry. Luciferase reporter gene assay was used to detect the transcriptional activity of MDR1 promoter. The results showed that TPL significantly decreased the resistance degree of K562/A02 cells, inhibited P-glycoprotein expression (mean fluorescent intensity decreased from 123 ± 13 to 39 ± 13) and increased the intracellular concentration of ADM (mean fluorescent intensity increased from 18 ± 5 to 34 ± 6) in K562/A02 cells. Luciferase reporter gene assay demonstrated that TPL inhibited the transcriptional activity of MDR1 promoter by 75%. It is concluded that TPL may effectively reverse the multidrug resistance in K562/A02 cells via modulating P-glycoprotein expression and increasing intracellular ADM accumulation.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Diterpenes ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Epoxy Compounds ; pharmacology ; Humans ; K562 Cells ; Phenanthrenes ; pharmacology ; Promoter Regions, Genetic

Objective To evaluate the capability and accuracy of multi-shce spiral computed tomography(MSCT)in detecting atherosclerotic plaques in nonstenotic coronary arteries with reference to the findings of intravascular ultrasound(IVUS)in a segment analysis.Methods Both IVUS exams and 16-row MSCT scans were performed on 35 consecutive patients among whom 30 patients had successful MSCT scans.A total of 94 coronary segments without significant coronary stenoses were paired-analyzed both on IVUS and MSCT segment by segment.The plaques were classified as calcified,fibrotic and soft types according to the echogeneity on IVUS.Plaque attenuation on MSCT was measured and expressed by Hounsfield units(HU).Results When referred to IVUS,MSCT had a sensitivity of 82.1%(46/56)and specificity of 89.5% (34/38),respectively in detectiong any plaques.For the detection of calcified plaques,the sensitivity and specificity were 92.1%(35/38)and 96.4%(54/56),respectively.For the detection of mixed and noncalcified plaques,MSCT had sensitivity of 73.2%(30/41)and specificity of 88.7%(47/53).But for the detection of the noncalcified plaque,the sensitivity was 66.7%(12/18). According to the findings On IVUS,the plaques were classified as calcified(n=19),fibrotic(n=19)and soft(n=16).The CT attenuation of calcified plaques was(489?169)HU(196 to 817 HU),fibrotic plaques(69?21)HU(25 to 117 HU)and soft plaques(23?18)HU(-12 to 47 HU).Nonparametric Kruskal-Wallis test revealed a significant difference of plaque attenuation among the three groups(P

OBJECTIVETo study early change features of microRNA (miRNA) in the peripheral blood of Sophorae Tonkinensis Radix et Rhizoma induced liver injury rats, and to look for the miRNA biomarkers in the peripheral blood of early liver injury.

METHODSSixty Wistar rats were randomly divided into the control group and the Sophorae Tonkinensis Radix et Rhizoma (abbreviated as STRR) group, 30 in each group. Rats in the STRR group was administered with STRR decoction at 12 g/kg (2 mL/100 g), while equal volume of the distilled water was given to those in the control group. Rats were anesthetized on day 3, 7, 14, and 28, and 28 days after withdrawal. The serum samples were withdrawn. The alanine aminotransferase (ALT), aspartate transaminase (AST), total bile (TBIL), alkaline phosphatase (ALP), total protein (TP), and albumin (ALB) were detected. The globulin (GLO) level was calculated. HE staining was performed on the liver tissue to observe the pathomorphological changes. The whole blood was collected on day 7, 14, and 28 to perform the microarray test. The differentially expressed miRNAs were screened and verified by RT-PCR.

RESULTSThe ALT activity obviously increased on day 7 - 28 in the STRR group (P <0.05). The histopathological results showed the degeneration and swelling of the liver cells on day 28. In the microarray test, there were 11, 22, and 13 up regulated expressed miRNAs on day 7, 14, and 28, respectively. There were 1, 13, 2 down regulated expressed miRNAs on day 7, 14, and 28, respectively. By target gene prediction and pathway analysis of differentially expressed miRNA on day 7, 14, and 28, they involved in regulating and controlling signal transduction, cellular interaction, cytoskeleton. Differentially expressed miRNA might possibly participate in the process of liver injury. The RT-PCR result of the expression of miR-291a-5p with the peak time efficiency on day 7 showed that the expressions of miR-291a-5p in the peripheral blood and the liver tissue were basically identical.

CONCLUSIONmiR-291a-5p could early indicate the liver injury, which could be taken as one of an early marker in STRR induced liver injury.

Animals ; Chemical and Drug Induced Liver Injury ; metabolism ; pathology ; Drugs, Chinese Herbal ; adverse effects ; Female ; Liver ; metabolism ; pathology ; Male ; MicroRNAs ; blood ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effect of water soluble extracts of traditional Chinese herbs on growth of mouse hair follicles and hair bulb cells in vitro.

METHODSMouse hair follicles and hair bulb cells were cultured in Williams E medium with (experimental groups) or without (control group) water soluble extracts of Chinese herbs; the experimental group was further divided into mixture and single herb groups. Hair growth was observed by microscopy and growth activity of hair bulb cells was detected by MTT colorimetric assay.

RESULTOn day 7 of culture, the hair growth in the mixture groups was faster than that in the control group (P<0.05). On day 3 and 5 of culture, the cell growth activity in the mixture groups was greater than that in the control group (P<0.05). While the hair growth and the cell growth activity between the single herb groups and the control group were not significantly different.

CONCLUSIONThe water soluble extracts of mixed traditional Chinese medicines can promote the growth of mouse hair in vitro and stimulate the proliferation of hair bulb cells; while those of the single traditional Chinese herb have no effect.

Angelica sinensis ; Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Hair ; drug effects ; growth & development ; Hair Cells, Auditory ; cytology ; drug effects ; Hair Follicle ; drug effects ; Mice ; Mice, Inbred C57BL ; Organ Culture Techniques