Background5-Nitro-2-(3-styrene-acrylic amine) benzoic acid ( NPPB),a chloride channel inhibitor,has a promoting effect on cell apoptosis in myocardial ischemia and reperfusion of domestic rabbit.The CIC chloride channel has been found in the ocular trabecular cells.However,the effect of NPPB on the shape and function of trabecular cells is unclear. Objective This study was performed to investigate the effect of NPPB on the proliferation,cell cycle progression and apoptosis of human trabecular meshwork cells.MethodsThe immortalized human trabcular cell strain was cultured,and logarithmic-phase cells were incubated in 96-well plates at a density of 1 ×106/ml.Different concentrations of NPPB (10,50,100 μ mol/L) were added to the medium,and the MTT assay was used to assess the growth and proliferation of the cells.Flow cytometry was used to evaluate the cell cycle.Then,100 mg/L 5-FU or 100 mg/L 5-FU + 100 μmol/L NPPB was used to induce cell apoptosis,which was assessed by Annexin V-PI.The membrane potential of mitochondria was examined using rhodamine 123 (△ψm).Results After 48 hours of treatment with NPPB,the abosorbency (A value) of the cells was gradually lowered with the increasing dose of NPPB,with significant differences among the 4 groups (F =7.230,P =0.006).Compared with the 10 μmol/L NPPB group,the A values were significantly declined in the 50 and 100 μmol/L NPPB groups (t =1.610,P =0.025 ;t =12.270,P =0.001 ).Forty-eight hours after exposure to NPPB,the percentage of cells in G0/G1 phase was increased and that in the S phase was decreased.The percentages of cells in different phases of cell cycle were significantly different in comparison with their control groups (without NPPB)( P＜0.05 ).Twenty-four and 48 hours after the treatment with 100 mg/L 5-FU,the apoptosis rates of the cells were raised in the 100 mg/L 5-FU group and 100 mg/L 5-FU + 100 μmol/L NPPB group compared to the without NPPB group (t24h =2.130,P =0.023;t48h =4.810,P=0.011 ) ;while that in the 100 mg/L 5-FU+100 μmol/L NPPB group was higher than the 100 mg/L 5-FU group ( t24 h =1.980,P =0.037 ; t48 h =1.290,P =0.028 ),and the mitochondrial membrane potential was lowered ( t24h =1.580,P =0.029 ; F48 h =6.200,P =0.015 ).Conclusions NPPB suppresses the proliferation of human trabecular cells and promotes the cells to enter S phase via the G1/S check point.In addition,ClC might be involved in an anti-apoptosis mechanism through the internal mitochondrial pathway.

Objective To observe changes of cognitive function and the expression of tumor necrosis factor alpha(TNF-α),interleukin 10(IL-10) in hippocampus of diabetic rats,and assess the role of inflammation in the possible pathogenesis of diabetic encephalopathy (DE).Methods 30 male SD rats were randomly divided into control group and diabetes mellitus group.After 4 weeks of feeding high fat diet,diabetes mellitus group according to 30mg/kg injected with streptozotocin to establish type 2 diabetic rat model.At the end of the experiment,cognition were evaluated using water maze test.The concentration of beta-amyloid(Aβ) in hippocampus of diabetic rats were detected through enzyme linked immunosorbent assay,and the expression of TNF-α,IL-10 were detected by Western blotting.The expression of Aβ,TNF-α,IL-10 were observed through immunohistochemistry.Results Time spent in the target quadrant in diabetes mellitus group was shorter than that in control group ((38.21± 3.68)s vs (42.10±2.62)s,t=3.105,P＜0.01).The frequency of crossing original platform site was less than that in control group((2.62±0.77) vs(3.69±0.95),t=3.184,P＜0.01).Compared with control group the expression of Aβ,TNF-α were higher(BothP＜0.01),and IL-10 were lower(P＜0.01)in diabetes mellitus group.The positive expression of Aβ,TNF-α were obviously and IL-10 were less obviously observed in diabetes mellitus group according to immunohistochemistry.Conclusion The cognitive decline in diabetic rats is possibly related to inflammatory cytokines expressing out of balance.

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Aquilaria sinensis can generate agarwood, which is closely related with endophyte. Up to now, studies mainly focused on the effects of endophytic fungi on agarwood formation, but studies about endophytic bacteria are rarely reported. In our research, the T-RFs and Shannon index of endophytic bacteria in samples of agarwood increase. The number of distinctive T-RFs fragments of corresponding samples in the same group accounted for more than 60% the number of total T-RFs fragments. In samples of no-agarwood, the dominant bacterial population are Anoxybacillus, Clostridium, Candidatus endobugula, Lysinibacillus. In samples of agarwood, the dominant bacterial population are Clostridium, Lysinibacillus, Luteimonas, phytoplasma. Besides, there are. specific T-RFs fragment in samples of agarwood and no-agarwood respectively. When we perform cluster analysis, we found samples of agarwood highly gather together and samples of no-agarwood highly gather together. This means community of endophytic bacteria emerge significant and regular changes during agarwood formation, which may be result of agarwood production, or maybe it is important reason of agarwood production. In this paper, we obtain more comprehensive and accurate community of endophytic bacteria in Aquilaria sinensis and it's variation during agarwood formation using T-RFLP, which is first study of effects of endophytic bacteria on agarwood formation, and will help to exploit resource of endophytic bacteria more reasonably.
Bacteria ; classification ; genetics ; isolation & purification ; Biodiversity ; Endophytes ; classification ; genetics ; isolation & purification ; Thymelaeaceae ; microbiology ; Wood ; microbiology

Objective To evaluate the b value of diffusion-weighted(DW)MRI in distinguishing between benign and malignant breast lesions.Methods Three diffusion-weighted sequences were implemented with 500,1000 and 2000 s/mm~2 b values respectively on 95 breast lesions in 83 patients.All lesions were confirmed by pathology.The apparent diffusion coefficient(ADC)values and signal intensity (SI)were recorded and compared in different lesions(breast cancer,benign lesion,cyst and normal beast tissue)with the same b value and the same lesions with the different b values.Results(1)The mean ADC value and SI of breast cancer were 1.375?0.378 and 839.713?360.493 respectively with b= 500 s/mm~2,1.176?0.311 and 459.314?229.609 with b=1000 s/mm~2,0.824?0.198 and 243.825? 110.616 with b=2000 s/mm~2.The differences in the mean ADC value were significant between two type lesions(cancer and benign lesion,cancer and cyst,cancer and normal breast tissue)with b values of 500 s/mm~2 and 1000 s/mm~2.But the significant differenee was only seen between cancer and benign lesions when b value was 2000 s/mm~2.(2)The one-side upper limits of 95% confidence interval of mean ADCs were adopted as the point to separate the malignant from the benign lesions,the sensitivity was 70.92%, 70.73% and 69.77%,the specificity was 77.19%,75.70% and 54.76%,the accuracy was 77.12%, 74.32% and 62.35% respectively with b values of 500 s/mm~2,1000 s/mm~2 and 2000 s/mm~2.The areas under ROC eurves were Az_(500)=0.775?0.046(P0.05).Conclusion DWI MRI is useful for the differential diagnosis of breast lesions with b values of 500 s/mm~2 and 1000 s/mm~2.

Objective To explore the roles of clara cell secretory protein(CCSP)and eosinophil cationic protein(ECP)in the pathoge-nesis of bronchial asthma and to evaluate their diagnostic value in asthmatic children.Methods Induced sputum samples were obtained from 31 asthmatic children during chronic persistent period and clinical remission period.According to global initiative for asthma(GINA),the total of 31 cases accepted systemic treatment by inhaling glucocorticoid.The patients included 18 boys and 13 girls aged from 3.7 to 12.0 years,and their average age was 7.6 years.Sputum CCSP concentrations were measured by enzyme-linked immunosorbent assay(ELISA).And the concentrations of sputum ECP were determined with Pharmacia UniCAP system.Results Asthmatic children had significantly lower CCSP levels in sputum during chronic persistent period compared with clinical remission period(P

Objective To explore roles of total immunoglobulin E(IgE),interleukin-13(IL-13) in asthmatic children,and relation-ship between IgE,IL-13 levels in serum and those in induced sputum.Methods Twenty-six children with asthma who were in chronic persistent period and 20 healthy children were enrolled.Serum and hypertonic saline-induced sputum were obtained in asthmatic children,and serum alone were obtained in control subjects.The levels of IgE were deteced in serum and induced sputum by Pharmacia UniCAP system,and levels of IL-13 were measured by enzyme linked immunosorbent assay(ELISA).Results Asthmatic children had significantly higher serum of IgE and IL-13 levels than those of healthy control group(P0.05).There was positive correlation of IL-13 in serum and induced sputum(r=0.432 P

The effect of C-terminal region residues on the substrate specificity of a novel cyclic imide hydrolase (CIH), a recombinant cyclic imide hydrolase (CIH293), and its mutants deleted or substituted at C-terminus (CIH291, CIH290, KK292-293EE) was reported. The substrate specificity and kinetic parameters of the mutants were analyzed by both the spectrophotometric assay and high-performance liquid chromatography. Results show that the substrate specificity of mutants was not obviously changed, but slightly low for the affinity between the substrate and enzyme, compared with the wild-type enzyme, CIH293. In conclusion, the last three residues of CIH293 play an important role for the enzyme activity.

ObjectiveTo investigate the incidence of skeletal and/or articular malformations in central core disease (CCD) patients, and their risk of malignant hyperthermia (MH) in orthopedics. Methods28 unrelated patients with central core disease collected in the past 23 years were analyzed retrospectively. Results22 patients showed limb muscle weakness, while 6 patients didn't show any clinical symptom. Skeletal and/or articular malformations appeared in 21 patients, including joint contracture in 11, joint dislocation in 9, scoliosis in 12, lordosis in 5 and thoracic deformity in 1. 6 of the 10 patients could not endure orthopedic operation because of MH attack or dubitable MH attack. Calcium-induced calcium release (CICR) test performed on other 8 patients with MH family history or MH attack showed all those patients were MH susceptibilities. ConclusionSkeletal and/or articular malformations are common in CCD, as well as MH attack in orthopedics.

Understanding the research methods for drug protein targets is crucial for the development of new drugs, clinical applications of drugs, drug mechanisms, and the pathogenesis of diseases. Cellular thermal shift assay (CETSA), a target research method without modification, has been widely used since its development. Now, there are various CETSA-based technology combinations, such as mass spectrometry-based cellular thermal shift assay (MS-CETSA), isothermal dose response-cellular thermal shift assay (ITDR-CETSA), amplified luminescent proximity homogeneous assay-cellular thermal shift assay (Alpha-CETSA), etc., which combine their respective advantages and further expand the application scope of CETSA. These technologies are suitable for the entire drug development chain, from drug screening to monitoring the target binding and off-target toxicity of drugs in patients. Based on the author's research experience, this paper reviews the principles of CETSA and related binding technologies, their application in target discovery, and the progress of data processing and analysis in recent years, aiming to provide reference and reference for the further application of CETSA.