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

Aged ; Humans ; Lower Extremity ; Middle Aged ; Muscle, Skeletal/physiology* ; Vibration

Background:Dexamethasone can protect mice against the acute inflammatory liver injury by inhibiting innate immune cell function. However,the roles of T cell in this protective effect remain unknown. Aims:To investigate the regulatory effect of dexamethasone on T cell immune response in acute inflammatory liver injury. Methods:Six male C57BL/ 6J mice were randomly divided into 2 groups. One hour before induction of acute inflammatory liver injury by lipopolysaccharide, dexamethasone 5 mg/ kg and PBS were given intraperitoneally in experimental group and model group,respectively. All the mice were sacrificed 12 hours after model construction. The clinical score and liver function parameters were assessed;splenic mononuclear cells were isolated for measurements of T cell activation,as well as cytokine expression,secretion, and transcriptional factor expression for different T-cell subsets. Results:Clinical score and serum levels of transaminase were significantly lower in experimental group when compared with the model group. Meanwhile,percentage of CD44 +CD62L - T cells(i. e. activated or memory T cells)from spleen was significantly decreased in experimental group. Among splenic T cell population,expression and secretion of IFN-γ,a Th1-type cytokine,was decreased;expression and secretion of IL-4,a Th2-type cytokine,percentage of regulatory T cells(Treg cells),and ratios of Th2 / Th1 and Treg/ Th1 were increased;transcriptional factor specific for Th1 cells was down-regulated,and those for Th2 and Treg cells were up-regulated. Conclusions:Dexamethasone inhibits T cell activation and directs the reciprocal T cell lineage differentiation (repressing Th1 cell differentiation,promoting Th2 and Treg cell differentiation),which may contribute to the protection against acute inflammatory liver injury.

Objective To investigate the synergistic effect of 11 β-hydroxysteriod dehydrogenase (11 β-HSD1) and S100A16 on the differentiation of3T3-L1 preadipocytes and its mechanism.Methods Lentiviral vectors PLJM1-11β-HSD1 and PLJM1-S100A16-GFP were respectively constructed and co-transfected into 3T3-L1 preadipocytes.The cell strains expressing 11 β-HSD1/S100A16 were screened with 2.5 μg/ml puromycin for two weeks.Western blot was employed to verify the lentiviral carrier transfection effects.The expressions of marker genes related to the adipocyte differentiation were detected by mean of realtime PCR.Oil red O staining was used to observe the lipid droplet accumulation and the content of triglyceride was measured after differentiation of preadipocytes.The effect of 11β-HSD1 and S100A16 on PPARγ promoter activity was detected by luciferase reporter gene.Results Compared with the empty vector group,the expressions of 11β-HSD1 and S100A16 protein in the lentivirus cotransfected 3T3-L1 cell strain were significantly higher.After 3T3-L1 cell strain co-expressing 1 1β-HSD1 and S100A16 was induced to differentiate for 8 days,the lipid droplets accumulation and triglyceride content were siginificantly increased,along with increased expressions of adipocyte differentiation marker genes such as PPARγ,CCAAT/enhancer binding protein α,lipoprotein lipase,fatty acid synthase,and adipocyte fatty acid-binding protein,in comparison with 11 β-HSD1 or S100A16 overexpression.The result of reporter gene indicated that 11 β-HSD1/ S100A16 enhanced PPARγ promoter activity.Conclusions 11β-HSD1 and S100A16 may jointly promote the differentiation of 3T3-L1 preadipocytes through a synergistic effect on PPARγexpression and play a critical role in the development of obesity.

OBJECTIVE Diabetes-induced endothelial cell (EC) dysfunction and neovasculariza-tion impairment constitute vascular complications with limited treatment regimens.Transcription factor FOXO1 is a key angiogenic regulator and plays a pathologic role in progression of diabetes.The pres-ent study was designed to determine the involvement of FOXO1 in impaired EC function and post-isch-emic neovascularization in diabetes and investigate underlying mechanisms.RESULTS We found that FOXO1-selective inhibitor AS1842856 improved blood flow recovery and capillary density in ischemic hindlimb,and rescued the delay of wound closure with a concomitant augmentation of mean perfusion rate in diabetic mice. In vitro,treatment with AS1842856 or FOXO1 siRNA abrogated high glucose-in-duced apoptosis and ameliorated capillary tube formation in human umbilical vein endothelial cells(HU-VECs). FOXO1 inhibition relieved alterations in mitochondrial networks and significantly suppressed the over production of mitochondrial reactive oxygen species(mtROS)induced by high glucose in ECs. Expression of dynamin-relatedprotein-1 (Drp1) and phosphorylation at Ser616, a protein required for mitochondrial fission, were enhanced by hyperglycemia, which could be neutralized by FOXO1 inhibition. Moreover, the transcription of Rho-associated coiled-coil containing protein kinase 1 (ROCK1), which phosphorylates Drp1 at Ser616, was shown by luciferase assay to be directly regulated by FOXO1. CONCLUSION These findings suggested that FOXO1 is critical to preserve mitochondrial quantity and func-tion in ECs,and FOXO1 may serve as a therapeutic target for microvascular complications of diabetes.

Objective To study effect of eleetromyograghie(EMG) biofeedback training on three kinds of u- rinary ineontifienee.Methods Nineteen patients with urinary incontinence were treated by means of EMG biofeed- back training twice a day for six weeks.The treatment was performed with a device,which can detect the EMG ampli- tude of the pelvic muscle and deliver electric stimulation accordingly.Results After 6 weeks of treatment,the inci- dence of uretbrorrhea was reduced by 41%,and the frequency of micturition was decreased by 38% ,while the fre- quency of urination in one day reduced to 9 to 13.The general subjectively rated improvement rate of patients was 53% ,while the general objectively one was 58%.Conclusion Biofeedback training has significant therapeutic: eftects on patients with urinary incontinence.

Objective To develop a rapid and efficient single cell PCR method for the non-invasive prenatal hemophilia A genetic diagnosis.Methods Six STR markers (DXS15,DXS9901,G6PD, DXS1073,DXS1108 and F8Civsl3) closely linked to F Ⅷ gene were examined in 118 healthy individuals and 12 hemophilia A families by muhi-plex fluorescent PCR.Three markers with the higher heterozygote rate and diagnostic rate were chosen and combined with amelogenin gene to establish the four-plex nested fluorescent PCR on single lymphocyte level.Results The single lymphocyte amplification rates of Amelogenin,DXS15,FSCivsl3 and DXSI073 were 94.3%,91.4%,100% and 100% in the normal male,and 100% ,97.1% ,97.1% and 97.1% in the normal female's respectively.The female was heterozygous in DXS1073 and DXS15.However,no allele dropout was found in both markers.Neither the false negative nor the false positive amplification was observed.Conclusion The single cell four-plex nested fluorescent PCR method established in this study is convenient and efficient,and will be hopefully employed in the clinical non-invasive prenatal genetic diagnosis for hemophilia A.

Objective To study the effectiveness and safety of lipo-PGE1 on proteinuria in patients with diabetic nephropathy. MethodsForty-eight cases suffering from diabetic nephropathy with proteinuria exceeding 0.5 g/d and stage Ⅱ-Ⅳ of chronic kidney disease were divided into two groups.The basic treatment scheme for all the patients includes insulin and/or oral antidiabetic drugs and anti-hypertension drugs.The patients in the control group were given dipyridamole,and those in the lipo-PGE1-treatment group were managed with lipo-PGE1(alprostadil injection),10 ?g,qd?14 d.Results After the treatment,a decrease in proteinuria and increase in serum albumin were observed in the lipo-PGE1-treatment group,which was superior to the control group(P

Objective To investigate the potential application of a non-viral gene carrier , TAT-LK15 , for delivering nNOSsiRNAin vivo and to study whether TAT-LK15/siRNA can be a new treatment method for chronic inflammatory pain. Method TAT-LK15 was complexed with nNOSsiRNA or scrambled control siRNA. The expression of nNOS was determined in SCDH of chronic inflammatory pain rats by western-blot assay. Pain control efficacy was evaluated by mechanical withdrawal threshold (MWT) and thermal withdrawal duration (TWD) assays. Results nNOS protein expression was efficiently inhibited by intrathecal injection of TAT-LK15/siRNA complexes , with the reduction of nNOS protein by 52%. Moreover , injection of TAT-LK15/siRNA com-plexes significantly could decrease MWT , but increase TWD in rats with chronic inflammatory pain. Conclusions TAT-LK15 can efficiently deliver nNOSsiRNAin vivo and nNOSsiRNA can relieve chronic inflammatory pain in rats.

Objective:To observe the effect of horse-riding squat exercise plus Governor Vessel-regulating Tuina(Chinese therapeutic massage)therapy on static balance function in patients with stroke.Methods:A total of 176 stroke patients were enrolled as the study subjects and divided into four groups by the random number table method,namely horse-riding squat+Tuina group,horse-riding squat group,Tuina group,and control group,with 44 cases in each group.The control group was treated with rehabilitation balance training.The horse-riding squat+Tuina group,horse-riding squat group,and Tuina group were treated with additional horse-riding squat exercise plus Tuina,horse-riding squat exercise,and Tuina treatment,respectively.Four weeks of treatment was regarded as one treatment course.After 1 treatment course,the balance function of the four groups was compared.Results:After treatment,the ellipse area of motion and the length of motion in all four groups were reduced,and the intra-group differences were all statistically significant(P＜0.05).The difference in the ellipse area of motion before and after treatment in the horse-riding squat+Tuina group and the horse-riding squat group was larger than that in the control group,and the differences were statistically significant(P＜0.05).The difference in the ellipse area of motion before and after treatment in the horse-riding squat+Tuina group was larger than that in the Tuina group,and the difference was statistically significant(P＜0.05).The difference in the length of motion before and after treatment in the horse-riding squat+Tuina group,the horse-riding squat group,and the Tuina group was greater than that in the control group,and the differences were statistically significant(P＜0.05).There was no statistically significant difference in the pairwise comparison between the horse-riding squat+Tuina group,the horse-riding squat group,and the Tuina group(P＞0.05).Conclusion:Horse-riding squat exercise plus Governor Vessel-regulating Tuina therapy can effectively improve the static balance function in patients with stroke.Training the affected lower limb weight-bearing exercise,adjusting the center of gravity distribution,and promoting the mutual balance and coordination between muscle groups may be the mechanism of improving the static balance function.