Objective To determine the effect of cheA gene of Helicobacter pylori in the bacterial chemotaxis in vitro and colonization in vivo. Methods The entire cheA and cheY genes were amplified and cloned from genomic DNA of H. pylori NCTC11637 strain. Subsequently, the prokaryotic expression systems of cheA and cheY genes were generated and the target recombinant proteins rCheA and rCheY were extracted by Ni-NTA affinity chromatography. Rabbits were immunized with either rCheA or rCheY for obtaining antisera, and rCheA-IgG and rCheY-IgG in the antisera were prepared using ammonium sulfate precipitation plus DEAE-52 column chromatography. A suicide plasmid of cheA gene was constructed and then a cheA gene knock-out mutant ( cheA - ) was generated based on homologous recombinant exchange using the suicide plasmid. The cheA- mutant was identified using PCR and sequencing. The phosphorylation levels of CheA and CheY molecules of cheA - and wild-type strain were determined by using rCheA-IgG and rCheY-IgG anchoring the target proteins and protein phosphorylation detection kit. The differences of chemotaxis in vitro and colonization in vivo between cheA- mutant and wild-type strain were compared using chemotactic model and BALB/c infection model of H. pylori. Results The cheA gene knock-out in genome of cheA- mutant was confirmed by the results of PCR and sequencing. After treated with 0. 001-0. 1 mol/L HCI for 10 min, the phosphorylation levels of CheA and CheY molecules of wild-type strain were rapidly descended from ( 59.6 ±11.5) μmol and (55.5 ± 10.2) μmol to ( 10.8 ± 2.6) and (5. 5 ± 1.2) μmol (P ＜ 0.05 ), while the phosphorylation of CheY molecule of cheA - mutant was no markedly changed with a persistent lower level ( P ＞0.05). The diameters [(10-20) ± (2-3) mm] of chemotactic aggregative rings of cheA- mutant were significantly less than those [(16-24) ± (2-3)mm] of wild-type strain (P ＜0.05). The positive isolation rate (90%) of H. pylori in gastric biopsy specimens of mice that infected with wild-type strain was remarkably higher than that (40%) of mice that infected with cheA- mutant (P ＜0.05). The result of fluorescence quantitative was also showed that the numbers (6.3 × 103 ±2.1 × 103 copies/mg) of H. pylori in gastric biopsy specimens of wild-type strain infected mice were significantly larger than those (8.3 × 101 ±3. 1 × 101 copies/mg) in gastric biopsy specimens ofcheA- mutant infected mice (P＜0.05). Conclusion The cheA gene of H. pylori has an important role in the bacterial chemotaxis in vitro and colonization in vivo.

A fast technique for implementing Holter cardio real time data reduction based on pointer and dual data buffers is presented in this paper, and an example of its application is also given here.
Algorithms ; Artificial Intelligence ; Computer Systems ; Data Compression ; methods ; Electrocardiography, Ambulatory ; instrumentation ; methods ; Equipment Design ; Humans ; Information Storage and Retrieval ; methods ; Signal Processing, Computer-Assisted ; Software

Alcoholism ; drug therapy ; metabolism ; physiopathology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hippocampus ; metabolism ; Learning ; drug effects ; physiology ; Male ; Memory ; drug effects ; physiology ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; genetics ; metabolism ; Superoxide Dismutase ; metabolism

Objective To investigate the roles and significances of MMP-2 and MMP-9 in diffuse proliferative lupus nephritis by repeated renal biopsy.Methods Seventeen patients diagnosed by renal biopsy as WHO typeⅣlupus nephritis were analyzed by immunohistochemistry staining for MMP-2 and MMP-9. Double staining for MMP-2 and MT1-MMP,MMP-9 and CD68 were also performed.Patients had repeated renal biopsy after followed up for 2.5 years.The relationship between expressions of gelatinases and pathological activity index and clinical data were studied.Results MMP-2 immunoreactivity was detected in normal controls and was increased in diffuse proliferative lupus nephritis.MMP-9 staining,which was almost negative in normal giomeruli,was increased much more significantly in diffuse proliferative lupus nephritis. The immunoreactivity of MMP-2 and MMP-9 was positive in MT1-MMP staining and CD68-positive macrophages, respectively.The expression of MMP-2 and MMP-9 was reduced by 70% and 62% in 10 patients whose clinical condition was partially alleviated,while the expressions in 7 patients whose clinical condition was not alleviated,were only reduced by 27% and 32%.The staining for MMP-2 and MMP-9 were correlated with activity index of lupus nephritis and proteinuria.Conclusion Up-regulation of gelatinases expression in diffuse proliferate lupus nephritis is correlated to activity index of the disease.

Fucoidan is one of the main bioactive components of polysaccharides. The current study was focused on the anti-tumor effects of fucoidan on human heptoma cell line HepG2 and the possible mechanisms. Fucoidan treatment resulted in cell cycle arrest and apoptosis of HepG2 cells in a dose-dependent manner detected by MTT assay, flow cytometry and fluorescent microscopy. The results of flow cytometric analysis revealed that fucoidan induced G2/M arrest in the cell cycle progression. Hoechst 33258 and Annexin V/PI staining results showed that the apoptotic cell number was increased, which was associated with a dose-dependent up-regulation of Bax and down-regulation of Bcl-2 and p-Stat3. In parallel, the up-regulation of p53 and the increase in reactive oxygen species were also observed, which may play important roles in the inhibition of HepG2 growth by fucoidan. In the meantime, Cyclin B1 and CDK1 were down-regulated by fucoidan treatment. Down-regulation of p-Stat3 by fucoidan resulted in apoptosis and an increase in ROS in response to fucoidan exposure. We therefore concluded that fucoidan induces apoptosis through the down-regulation of p-Stat3. These results suggest that fucoidan may be used as a novel anti-cancer agent for hepatocarcinoma.

Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species (ROS) levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.

y screen and prevent CAD in these people before diabetes sets in.

Background Q-value is used to express the asphricity of the anterior corneal surface.As a optical surface,the optial morphology of cornea is composed of a series of tangential sections that cut-off through the optical axial.Therefore,tangential section could represent the true optical morphology of the corneal surface,and Q-value calculation by tangential radius can better represent the asphericity.Objective This study was to calculate Q-value of meridian section of the anterior corneal surface by tangential radius of the tangential topography and analyze the corneal asphericity in horizontal interval.Methods Ninety right eyes of 90 myopia subjects aged 16-30years and with mean spherical equivalent of (-5.45 ± 2.75) D received corneal topography examination using Orbscan Ⅱ system.The subjects were assigned to low myopia group,moderate myopia group and high myopia group based on their diopter and 30 eyes for each group.The tangential curvatures on meridian section at a 0.1 mm interval from apex to periphery of the anterior surface were accepted and the Q value of the semimeridian section was calculated by linear regression mathematical formulas of the tangential radius.Mydriatic optometry,intraocular pressure,keratometer and fundus examinations were performed on the subjects.Written informed consent was obtained from each subject before any medical examination.Results The coefficients of determination in all the semimeridians were over 0.5.The average calculated Q-values in the nasal and temporal horizontal interval were -0.32±0.11 and-0.30±0.12,with a significant difference between them (t =2.009,P＜0.05).The vertex radius of curvature was (7.78±0.27)mm and (7.72±0.25)mm respectively in the nasal and temporal horizontal interval,showing a significant difference(t=-1.016,P＞0.05).No significant difference was seen in Q values of both nasal and temporal areas among three myopic groups (nasal:F =0.192,P =0.825 ; temporal:F =0.912,P =0.406).The average Q value of the nasal and temporal principal meridian was-0.33±0.14 and-0.30±0.13 respectively,and the r0 was 7.76±0.30 and 7.74±0.24 respectively.A weak positive correlation was found between r0 and mean Q value of nasal meridian (r=0.320,P＜0.05),but no significant correlation was found between r0 and mean Q-value of temporal meridian (r=0.104,P＞0.05).No significant differenees were seen in the Q values between nasal meridian and nasal zone (t=0.349,P＞0.05) as well as between temporal meridian and temporal zone(t=-0.373,P＞0.05).Conclusions The study analyzes the calculated Q-value of the semimeridian section in borizontal area with myopia by linear regression mathematical formulas of tangential radius on tangential topography.The anterior surface of the cornea is proved to be prolate ellipse in shape in the subjects with myopia.

Background Selective laser trabeculoplasty(SLT)has been widely accepted for the treatment of open-angle glaucoma.However,there are few reports about its application in treating angle-closure glaucoma so far.Objective This study was to clinically evaluate the safety and efficacy of selective laser trabeculoplasty for primary angle-closure glaucoma with opened anterior angle follow-up surgery.Methods A case-observational design was adopted in this study.A total of 33 eyes from 23 angle-closure glaucomatous patients with opened 180° angle were enrolled.All the patients understood this trial and written informed consent was obtained prior to the medical treatment.The average preoperative intraocular pressure(IOP)and number of medication were(23.07±4.01)mmHg and 1.52±0.66,respectively.A frequency doubled Q-switched Nd:YAG laser was used to irradiate the trabecular meshwork with 70-80 laser spots over 180°.The same preoperative anti-glaucoma drugs were topically administered during the follow-up period.Visual acuity,IOP,slit lamp examination,gonioscopy and the coefficient of outflow facility were regularly assessed and performed after laser surgery.Results The IOP values of the 33 eyes were(18.78±4.87)mmHg,(18.83±3.95)mmHg,(19.78±4.48)mmHg,(19.63±3.68)mmHg,(19.11±3.36)mmHg,and(21.17±3.14)mmHg 1 day,l week,and 1,3,6,9 months after operation,showing significant decline in comparison with preoperation(P＜0.01).The mean drug uses were 1.63±0.85,1.23±1.01,1.25±0.98,1.21±1.02,1.22±0.86,and 1.69±0.53 at the same time point after operation,respectively,and significant differences were found 1 week,1,3,6 months in postoperation in comparison with preoperation(P =0.020,0.026,0.020,0.031).Ciliary congestion and aqueous flare naturally disappeared 1 week after surgery.The coefficients of outflow facility were 0.11±0.09 and 0.23±0.15,respectively at pretreatment and 3 months after SLT with a statistically significant difference (P=0.002).Conclusions Selective laser trabeculoplasty is effective and safe for angle-closure glaucoma.However,it is essential to have 1/2 circle of anterior chamber open for SLT in angle-closure glaucoma.

The traditional conotation of combination of disease and syndrome means the combination of disease differentiation in traditional Chinese medicine (TCM) diseases and TCM syndrome differentiation,which was derived from the Yellow Emperor's Inner Classic.And the Treatise on Cold Damage and Miscellaneous Diseases describes the clinical application of this theory.Under the background of integrative medicine,the modern conotation of combination of disease and syndrome means TCM syndrome differentiation and treatment based on modern medicine disease differentiation,which is commonly used in the current clinical practice.The disease and syndrome combination diagnosis mode based on syndrome,etiology,pathology and pharmacology can understand the disease from a multiple and comprehensive level,in order to realize the combination and complementation of advantages from both medicines.In addition,formula-syndrome relation means the relationship between the formula and its relevant indications.In general,indications often involve syndromes,symptoms,diseases and body constitutions.And the application of formula in the Treatise on Cold Damage and Miscellaneous Diseases is the representative of this theory.Hence,the application of associated classical formula gives example to the clinical formula-syndrome relation application in modem times.