Objective To investigate the role of Th17 cells and related cytokines in Helicobacter pylori(H.pylori) infection.Methods To establish a mouse model of H.pyloriinfectious gastritis.Meanwhile,a treatment group and control group were set up.The histological changes in the gastric mucosa were observed and scored under light microscopy;The expression of IL-17 and IL-23 mRNAs as well as protein in gastric tissue was detected by RT-PCR and ELISA,respectively;Th17 cells in mice splenocyte were evaluated using the flow cytometry (FCM) method.Results We found significantly higher levels of IL-17 and IL-23 protein and mRNA levels in supernatant of gastric tissue homogenates of infection group as compared to controls and Th17 cells in spleen from mice of H.pylori infection group were all obviously higher than that of control group;IL-17 mRNA,IL-23 mRNA and the ratio of Th17 cells in mice splenocyte of H.pylori infection group mice increased gradually with the time prolonged;The levels of both mRNA and protein levels of IL-17and IL-23 decreased significantly in the treatment group compared with pre-treatment;There was a positive correlation between IL-17 and IL-23 expression levels of mice gastric mucosa and gastric inflammation degree.Conclusion H.pylori infection induced the immune response of Th17 cells;The levels of IL-17 and IL-23 increased in gastric mucosa of mice after H.pyloriinfection;The degree of H.pylori-infected mice gastric inflammation was positive correlation with the levels of IL-17 and IL-23 of gastric mucosa.

Background Subretinal transplantation of retinal pigment epithelium (RPE) cells for the treatment of age-related macular degeneration (AMD) have accelerated the drive to develop xeno-free cultivation system that support the rapid differentiation of human embryonic stem cells (hESCs) into ES-RPE cells.Objective This study was to report a modified xeno-free culture system and method for accelerating derivation of hESCs to differentiate into RPE cells.Methods This study was approved by Ethic Committee of Zhongshan Ophthalmic Center.HESC H1 line was cloned and cuhured in Vitronectin XFTM-coated 6-well dish with xenogenetic-free medium.Cells were cultured in 50 ng/ml noggin,10 ng/ml DKK-1 and 10 ng/ml insulin like growth factor-1 (IGF-1) medium for 2 days,and then the concentration of noggin was decreased to 10 ng/ml and 5 ng/ml basic fibroblast growth factor (bFGF) and cultured for the following 2 days.Sequentially,noggin and bFGF were removed and cultured for 2 days.Finally,1 μmol/L CHIR99021 was added in medium for 6 days.Morphological changes in the progress of ESCs differentiation into RPE were observed by Living Cell Imaging System.The expression of Mitf and RPE65,RPE cellsspecific markers,in the cells were detected by immunofluorescence technique,and the relative expression levels of RPE cells-specific marker mRNA were assayed using real time fluorescent quantitation PCR.Results Polygonalshape monolayer cells which contained pigments were initially observed at day 14 after cultured with the cobblestonelike arrangement.Mitf and RPE65 were strongly expressed in the hES-derived RPE cells 35 days after induced,showing red fluorescence,and the cells presented hexagonal shape at cultured day 60 with numerous pigment granules in cytoplasm.Compared with before differentiation,the expression levels of Mitf mRNA in hES-RPE cells increased by (3.43±2.77) folds and (8.91 ± 2.83) folds,and the expression levels of RPE65 mRNA increased by (14.60 ± 3.94) folds and (87.16 ±9.32) folds at day 7 and day 14 after differentiation,respectively (all at P＜0.05).Conclusions A defined xeno-free culture system is successfully established by adding niacinamide,DKK-l,noggin,IGF-1 and CHIR99021 in xeno-free medium,and this system can accelerate the derivation and differentiation of hESCs into RPE-like cells.

Objective: To observe clinical efficacy of oral folic acid (FA) intervene in hyper-homocysteinemia (HHcy) patients combining coronary artery disease (CAD) and heart failure (HF), to study the effect of blood level of Hcy on cardiac function. Methods: A total of 126 relevant patients with blood level of Hcy>15 μmol/L were randomly divided into 2 groups:Routine group, the patients received anti-platelet therapy, statins, beta-blockers, diuretics, angiotensin converting enzyme inhibitor (ACEI) or angiotensin II receptor antagonist and FA group, in addition to above mentioned therapies, the patients also received FA 5 mg/day. n=63 in each group and all patients were treated for 3 months. Fasting blood levels of Hcy, BNP and left ventricular end diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF), 6-minute walk test (6MWT) were compared between 2 groups at pre- and 3 months post-treatment. Results: ① Based on NYHA classification, the patients with cardiac function at II, III, IV had accordingly increased blood levels of Hcy, BNP and LVEDD, while decreased LVEF and 6MWT, all P<0.05. ② Blood levels of Hcy were positively related to BNP (r=0.733, P<0.001) and LVEDD (r=0.511, P<0.001), negatively related to LVEF (r=-0.382, P<0.001) and 6MWT (r=-0.410, P<0.001). ③ With 3 months treatment, FA group and Routine group showed decreased Hcy level as (8.43 ± 1.87) μmol/L vs (3.29 ±1.68) μmol/L and BNP (891.84 ± 456.10) pg/ml vs (682.24 ± 463.79) pg/ml, reduced LVEDD (4.33 ± 1.231) mm vs (2.06 ± 1.73) mm, while elevated LVEF (6.59 ± 2.28) % vs (2.52 ± 2.37) % and 6MWT (142.97 ± 55.15) m vs (86.35 ± 59.06) m, all P<0.05. Conclusion: Increased blood level of Hcy is risky for HF occurrence, FA may treat HHcy and further improve the cardiac structure and function in HF patients.

OBJECTIVETo explore the protective mechanism of electroacupuncture (EA) at "Neiguan" (PC 6) on ischemic myocardial injury, and to explain the response patterns and characteristics of the specific effect of acupoints along meridians in sodium channel in the level of cardiac organ.

METHODSA total of 60 SPF male rats were randomly divided into a blank group, a model group, a non-acupoint group, a Neiguan group and a Lieque group, 12 cases in each one. Except the blank group, rats in the remaining group were treated with subcutaneous injection of isoprenaline to establish the model of myocardial ischemia. Rats in the Neiguan group, Lieque group and non- acupoint group were treated with EA, dilatational wave, with a frequency of 2 Hz/20 Hz. The intensity was 2-3 mA. The needles were retained for 20 min per time, once a day for consecutive 7 days. In the blank group and control group, the rats were grasped and fixed at the treating time each day. The western-blot method was used to test the expression of voltage-gated sodium channel alpha subunit (Nav 1.5), protein tyrosine kinase (PTKs) and protein tyrosine phosphatase (PTPs).

RESULTSThe expression of Nav 1.5 and PTKs in the model group was lower than that in the blank group (both P<0. 01); the expression in the Neiguan group and Lieque group was higher than that in the model group (all P < 0.01); the expression of Nav 1.5 and PTKs in the Neiguan group was higher than that in the Lieque group (both P < 0.01). The expression of PTPs in the model group and non-acupoint group was higher than that in the blank group (both P < 0.01); the expression of PTPs in the Neiguan group and Lieque group was significantly down-regulated, which was lower than the model group (both P < 0.01); the down-regulation in the Neiguan group was significantly different from that in the Lieque group (P < 0.05).

CONCLUSIONEA at "Neiguan" (PC 6), by down-regulating the expression of PTPs, up-regulating the expression of Nav 1.5 and PTKs, is likely to achieve the aim of regulation on sodium channel activity and calcium overload, further to improve myocardial ischemia, which provides experimental basis for the theory of the specific effect of acupoints along meridians.

Acupuncture Points ; Animals ; Disease Models, Animal ; Electroacupuncture ; Humans ; Male ; Myocardial Ischemia ; genetics ; metabolism ; therapy ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium Channels ; genetics ; metabolism

Objective The staffs of biosafety level 3 laboratories (BSL-3) face with the stress of handling highly pathogenic microbs and special laboratory environment.The job stress may result in accidents in the laboratory as negative factor for the risk control.The research may provide support for the control of risk in biosafety laboratories.Methods In order to assess the job stress in the staff in BSL-3 laboratory, we modified “the Chinese simple job stress questionnaire”based on the theory of the JDC mode and ERI mode, and an investigation was carried out.The present study included the staffs (87 employees) from six BSL-3 laboratories located in five provinces ( Shanghai, Zhejiang, Jiangsu, Fujian and Wuhan) .Results Analysis of the data indicates that variables of age, working years, job duties, manipulating of animals, type of microorganisms and transmission route have a significant influence on the level of job stress in BSL-3 laboratory.Conclusion The BSL-3 laboratory staff in higher stress level have the characteristicses:20-39 years old, short work years, regular staff, operating on air-borne microbiology, manipulating of animals and operating on one more microbiology.

Reduced sulfur compounds ( RSCs) are one of the main pollutant species in the atmosphere, so it is of great significance to develop a rapid and on-line approaches for their detection. In this study, a portable time-of-flight mass spectrometer ( TOF-MS) with magnetic field enhanced photoelectron ionization source was designed to detect RSCs. The photoelectron ionization source was induced from vacuum ultraviolet photons which generated from vacuum ultraviolet (VUV) lamp with energy of 10. 6 eV. The energy of photoelectrons was controlled by adjusting the extraction voltage to produce the photoelectron ionization, and an annular magnet was used in the ionization region to improve the ionization efficiency of photoelectrons. From the simulation result by SIMION software, it was found that the introduction of magnet field made the motion trajectroies of electrons in the helical motion increase and the convergence of electron at the ionization source was achieved. Experimental results showed that after introducing the magnet filed, the sensitivity of H2 S, SO2 and CS2 was improved by a factor of 5. 3, 9. 4 and 6. 9, respectively. With a detection time of 50 s, the limits of detection for H2S, SO2 and CS2 were 0. 14, 0. 52 and 0. 31 mg/m3(S/N=3), respectively. It could be concluded that the portable TOF-MS with magnetic field enhanced photoelectron ionization source has great potential to be applied for on-line monitoring of volatile sulfides at the emission source.

Objective: To investigate the intervention effect of SB431542, which inhibits the TGF-β/Smad3 signaling pathway, on silicotic fibrosis in rats. Methods: A total of 40 specific pathogen-free Sprague-Dawley rats were divided into normal saline control group, model group, SB431542 inhibitor group, and SB431542 inhibitor control group using a random number table, with 10 rats in each group. All rats except those in the normal saline control group were given non-exposed single intratracheal instillation of free silicon dioxide dust suspension 1 mL (50 mg/mL) ; the rats in the SB431542 inhibitor group were given intraperitoneal injection of SB431542 (5 mg/kg) on days 7 and 30 after dust exposure, those in the SB431542 inhibitor control group were given intraperitoneal injection of SB431542 cosolvent (5 mg/kg) on days 7 and 30 after dust exposure, and those in the normal saline control group were given intratracheal instillation of an equal volume of normal saline (5 mg/kg). On day 60 after dust exposure, the paraffin-embedded section of the right upper lobe of lung was collected for HE staining; the left upper lobe of lung was collected to measure the mRNA levels of fibronectin (FN) , collagen type I (COL-I) , and collagen type III (COL-III) by quantitative real-time PCR; the right inferior lobe of lung was collected to measure the protein levels of FN, COL-I, COL-III, phosphorylated Smad3 (p-Smad3) , and Smad3. Results: Compared with the normal saline control group, the model group had nodules with various sizes in lung tissue, with rupture of some alveolar septa, emphysema changes, and pulmonary interstitial fibrosis, as well as significant increases in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 in lung tissue (P<0.05) . Compared with the SB431542 inhibitor control group, the SB431542 inhibitor group had a relatively complete structure of lung tissue without marked nodules and with a small amount of exudate in alveolar space and the lumen of bronchioles, as well as significant reductions in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 in lung tissue (P<0.05) . There were no significant differences in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 between the model group and the SB431542 inhibitor control group (P>0.05) . Conclusion SB431542 exerts an intervention effect on silicotic fibrosis by blocking the TGF-β/Smad3 signaling pathway and reducing the expression of the downstream fibrosis factors FN, COL-I, and COL-III.

The aim of this study was to investigate the effects of tyrosine kinase inhibitor PTK787 on cell proliferation, cell cycle and the expression of fak mRNA of human chronic myeloid leukemia (CML) cell line K562, and to explore the mechanism of PTK787 against acute myeloid leukemia. The MTT method was used to detect the effects of PTK787 in various concentrations and at different time points on proliferation of K562 cells; the flow cytometry was used to determine the effects of PTK787 in different concentrations on cell cycle of K562 cells; the RT-PCR was used to assay the expression of fak mRNA in K562 cells treated with PTK787 for 48 hours. The results showed that along with increasing of the concentration and prolonging of time, the inhibitory rate of PTK787 on K562 proliferation was gradually enhanced. The comparison between various concentration groups at same time or comparison between various time groups in same concentration showed significant differences (p < 0.05), in which the effect of 320 micromol/L PTK787 on cells was strongest, while the continuous increase of PTK787 concentration or prolong of action time did not enhance the inhibitory rate on K562 proliferation. With increasing of drug concentration, the cell proportion in G(1) phase gradually increased, the cell proportion in S phase gradually decreased, the comparison between various groups revealed significant differences (p < 0.05), however the continuous increase of drug concentration from 160 micromol/L did not obviously change the cell proportion in phases of cell cycle. With increasing of drug concentration, the expression of fak mRNA in K562 cells gradually reduced with significant differences between various groups (p < 0.05), but with continuous increase of drug concentration from 160 micromol/L, the effect of PTK787 on the expression of fak mRNA in K562 cells also did not obviously change. It is concluded that the PTK787 shows effect of anti-leukemia cells through inhibiting transformation of the K562 cells from G(1) phase into S phase and decreasing the expression of fak mRNA in cells.
Cell Cycle ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 1 ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Phthalazines ; pharmacology ; Pyridines ; pharmacology ; RNA, Messenger ; genetics

This study is to investigate the effects of concentration, intestinal section, pH, paracellular route, substrate/inhibitor of enzyme (CYP3A) and proteins (P-gp, MRP2, SGL1) on the absorption of forsythoside A. The absorption of three concentrations (2.6, 5.2, and 10.4 microg x mL(-1)) of forsythoside A in different intestinal segments was studied with phenol red as the marker by rat circulation in situ. The results showed that the residue of forsythoside A with different concentrations had little significant difference from that obtained after perfusing via duodenum, jejunum, ileum and colon, which indicated that the absorption of forsythoside A was passive diffusion and had no difference in different segments of rat intestine. The residue of forsythoside A increased to 466.160 and 463.429 microg respectively when cyclosporine (4 microg x mL(-1)) or midazolam (50 micromol x L(-1)) was added to the circulation fluid, which showed significant difference compared to the control group (P < 0.05). Moreover, the residue of forsythoside A showed a tendency of increase with the increase of cyclosporine or midazolam. When digoxin (50 micromol x L(-1)) or EDTA (10 microg x mL(-1)) was added to the circulation fluid, the residue of forsythoside A decreased to 325.110 and 369.888 microg respectively, which showed significant difference as compared to the control group (P < 0.05). Besides, the residue of forsythoside A showed a tendency of reduction with the increase of digoxin or EDTA. However, there is no significant change in the absorption of forsythoside A when the different concentrations of mannitol were added to the circulation fluid. The results above indicated that the absorption of forsythoside A was mainly passive diffusion and involved paracellular route at the same time. In addition, the substrates of P-gp or CYP3A had dose-dependent effect on the absorption of forsythoside A.
Animals ; Colon ; metabolism ; Cyclosporine ; pharmacology ; Digoxin ; pharmacology ; Dose-Response Relationship, Drug ; Duodenum ; metabolism ; Edetic Acid ; pharmacology ; Glycosides ; administration & dosage ; pharmacokinetics ; Hydrogen-Ion Concentration ; Ileum ; metabolism ; Intestinal Absorption ; Jejunum ; metabolism ; Male ; Mannitol ; pharmacology ; Midazolam ; pharmacology ; Rats ; Rats, Sprague-Dawley