2.Regulation of nitric oxide donor JS-K on tumor energy metabolism in H22 tumor-bearing mice
LIU LING ; HUANG ZI-LE ; WANG JIAN-GANG
Chinese Journal of Pharmacology and Toxicology 2017;31(10):964-965
OBJECTIVE To investigate the regulation of {O2 (2,4-dinitrophenyl)1-〔(4-ethoxycarbonyl) piperazin-1-yl〕diazen-1-ium-1,2-diolate}(JS-K), anitric oxide donor, on tumor energy metabolism in H22 tumor- bearing mice. METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line. The JS-K group and model group were received JS-K (0.75 and 1.5 mg?kg-1) and saline via tail intravenous once every 3 d for 14 d, received 5 injections, respectively. The positive group was received 5-FU 20 mg·kg- 1 by intraperitoneal injection once a day for 14 d. On the 15th day mice were sacrificed. The tumor growth inhibition rate were calculated. The activities of hexokinase (HK), phospho?fructo kinase (PFK), pyruvate kinase (PK), succinate dehydrogenase (SDH), adenosine triphosphatase (ATPase), and the levels of lactic acid (LD) and adenosine triphosphate (ATP) in tumor tissues were de?termined by colorimetric method. RESULTS Compared with model group, the tumor mass of JS- K 0.75 and 1.5 mg·kg- 1group was significantly reduced (P<0.01),and the tumor growth inhibition rate was 23.9% and 50.3%, respectively. The activity of HK, PFK, PK, SDH and ATPase of tumor tissue in model group was (22.6±3.7, 14.4±2.6, 12.9±3.2 and 10.5±2.6)U·g-1 protein and (0.70±0.10)μmolPi·mg-1 protein per hour, respectively; which in JS-K 1.5 mg?kg-1 group was dropped by 42.0%, 26.6%, 22.7%, 23.3% and 21.7% (P<0.01, P<0.05). Compared with the model group, the level of ATP and LD in JS-K group was dropped (P<0.01). CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism with inhibition of glycolysis and aerobic oxidation.
3. Effects of aqueous extract of Polygoni Multiflori Radix and its main constituents on expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells
Chinese Traditional and Herbal Drugs 2017;48(23):4912-4920
Objective To analyze the main chemical constituents and their contents in aqueous extract of Polygoni Multiflori Radix (PMR, root of Polygonum multiflorum), and to elucidate the effects of aqueous extract of PMR and its main constituents on the expression of the mRNA of CYP1A2, CYP2C9, and CYP2E1 in human liver L02 cells. Methods The main chemical constituents and their content in aqueous extract of PMR were determined by HPLC. The cytotoxicity of aqueous extract of PMR and its main constituents on L02 cells was determined by MTT assay. The mRNA expression of CYP1A2, CYP2C9, and CYP2E1 in L02 cells were detected by quantitative real-time PCR. Results There were four main well-separated chromatographic peaks standing for tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside, emodin and physcion in aqueous extract of PMR. The content of thesecomponents in aqueous extract of PMR was (1.14 ± 0.03)%, (0.106 9 ± 0.001 6)%, (0.010 8 ± 0.000 9)%, (0.003 55 ± 0.000 19)%, respectively. The cytotoxicity of aqueous extract of PMR and emodin on L02 cells at 24 h was dose-dependent, and the concentration of 50% inhibition was 7.290 mg/mL and 0.082 mmol/L respectively. Tetrahydroxy stilbene glucoside, emodin-8-O-β-D-glucoside and physcion did not show significant cytotoxicity on L02 cells in the experimental concentrations. Aqueous extract of PMR and emodin significantly inhibited the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells. Emodin-8-O-β-D-glucoside inhibited the expression of mRNA of CYP1A2 and CYP2C9. Tetrahydroxy stilbene glucoside inhibited the expression of mRNA of CYP1A2 but activated the expression of mRNA of CYP2C9. Physcion inhibited the expression of mRNA of CYP1A2 and CYP2C9 in a dose-dependent manner, but inhibited the expression of mRNA of CYP2E1 in low concentration and activitated the expression of mRNA of CYP2E1 in high concentration. Conclusion The inhibition of aqueous extract of PMR on the expression of mRNA of CYP1A2, CYP2C9, and CYP2E1 in L02 cells is the combined effect of all components in it. The main four components all inhibit the expression of mRNA of CYP1A2. The anthraquinone is the main component inhibiting the expression of mRNA of CYP2C9. The free anthraquinone is the main component inhibiting the expression of mRNA of CYP2E1.
4.The Structural Parameters-functional Activity Relationship of Alpha-helical Antimicrobial Peptides
Tian-Tang DONG ; Zi-Gang TIAN ; Jian-Hua WANG ;
China Biotechnology 2006;0(09):-
In response to problems from the development of multi-drug-resistant pathogenic bacteria,it is urgent to find new antimicrobials.Antimicrobial peptides(AMPs) are a kind of ideal new antimicrobials with the advantages including their potential for broad-spectrum activity,rapid bactericidal activity and low propensity for resistance development,and have a bright future.Alpha-helical antimicrobial peptides are a main kind of AMPs.The following features was reviewed and elucidated including the structure and activity relationship(SAR) from different aspects including the degree of helicity,hydrophobic moment,hydrophobicity,net positive charges and so on,and the application of SAR on the molecular design and improvement of AMPs.
5.Bacteriophage Lysins:A Novel Effective Antibacterial Agents
Yuan-Zi FANG ; Yan WANG ; Jian-He SUN ;
Microbiology 2008;0(12):-
Lysins are efficient bacteria cell wall digesting enzymes encoded by DNA bacteriophage. Gram-positive bacteriophage lysins feature similar domain structure, high lytic efficiency, synergic antibacterial effect with antibiotics, rare neutralization by antibodies, less chance of developing drug-resistant strains, et al. The past decade has seen a considerable amount of research worldwidely focused on lysin, and lysins have been used successfully in a variety of animal models to control pathogenic antibiotic resistant bacteria found on mucosal surfaces and infected tissues. The great potential of lysins as an anti-infective agent prompted this review.
6.Clinical Study of Postoperation Complications in 100 Patients with Supratentorial Cerebral Hemisphere Gliomas
Zi WANG ; Jian XIE ; Tao JIANG ; Song PU
Journal of Medical Research 2006;0(01):-
Objective To study the condition of postoperation complications in patients with supratentorial cerebral hemisphere gliomas,and to provide objective references for clinical classification and treatment. Methods Data of 100 patients with supratentorial cerebral hemisphere gliomas,treated surgicaly in the department of neurosurgery,gliomas center of Tiantan hospital from June 2006 to December 2007,were reviewed and analyzed.We classified postoperative complications as neurological,regional and systemic complications,and especially mainly analyzed the reasons of oceching,prevention and cure of the neurological postoperative complications.Results Through the complete preoperative evaluation in patients to choose the proper operative route,methods and technique means can improve postoperative KPS score compare with preoperative obviously ,which can achieve better operation result.Conclusion Neurosurgeon must design consummately individualization operation plan depending on patient history,neurology symptoms,physical sign,and preoperative check,and understand the operation anatomy including the pathology anatomy and function anatomy. They must foresee the postoperative complications,prevent it as far as possible,and treat it reasonable in time.
8.A novel microtiter plate radioimmunoassay of insulin autoantibody
Can, HUANC ; Zhang-wei, LI ; He-lai, JIN ; Xia, WANG ; Jian-ping, WANG ; Zi-guang, ZHUO
Chinese Journal of Nuclear Medicine 2009;29(1):50-54
Objective Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel microtiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods Diluted 125Ⅰ-insulin was mixed with 5 ul serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4℃). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1 DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, x2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results (1) The optimized testing condition was described as 2×104 CPM of 125Ⅰ-insulin, 5 ul serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9 %(Kappa value=0.402). There were 25 samples with discordant results, which were positive for IAA titer using the corresponding microtiter plate RIA but negative using the novel RIA kit. (3) In TIDM group the positive rate of IAA was 19.7% (16/71), higher than the healthy controls (0.9%, x2=54.36, P<0.001). The subgroup of T1DM children (with 0-9 years) showed the highest IAA positive rate (55.6% ,x2=4.85, P<0.05). In T2DM group the frequency of IAA was 1.5% (8/551), which had no significant difference comparing with that of healthy controls (x2= 0.95, P >0.05). Conclusions Our proposed microtiter plate RIA method for IAA is highly sensitive and specific, likely to be feasible for clinical application. The frequency of IAA is high in children with T1DM.
9.Characterization of Synonymous Codon Usage Bias in the Pseudorabies Virus US1 Gene
Meili LI ; Zhiyao ZHAO ; Jianhong CHEN ; Bingyun WANG ; Zi LI ; Jian LI ; Mingsheng CAI
Virologica Sinica 2012;27(5):303-315
In the present study,we examined the codon usage bias between pseudorabies virus (PRV) US1 gene and the US1-like genes of 20 reference alphaherpesviruses.Comparative analysis showed noticeable disparities of the synonymous codon usage bias in the 21 alphaherpesviruses,indicated by codon adaptation index,effective number of codons (ENc) and GC3s value.The codon usage pattern of PRV US1 gene was phylogenetically conserved and similar to that of the US1-like genes of the genus Varicellovirus of alphaherpesvirus,with a strong bias towards the codons with C and G at the third codon position.Cluster analysis of codon usage pattern of PRV US1 gene with its reference alphaherpesviruses demonstrated that the codon usage bias of US1-like genes of 21 alphaherpesviruses had a very close relation with their gene functions.ENc-plot revealed that the genetic heterogeneity in PRV US1 gene and the 20 reference alphaherpesviruses was constrained by G+C content,as well as the gene length.In addition,comparison of codon preferences in the US1 gene of PRV with those of E.coli,yeast and human revealed that there were 50 codons showing distinct usage differences between PRV and yeast,49 between PRV and human,but 48 between PRV and E.coli.Although there were slightly fewer differences in codon usages between E.coli and PRV,the difference is unlikely to be statistically significant,and experimental studies are necessary to establish the most suitable expression system for PRV US1.In conclusion,these results may improve our understanding of the evolution,pathogenesis and functional studies of PRV,as well as contributing to the area of herpesvirus research or even studies with other viruses.