Investigation on the potential adverse outcome pathway of the sensitive endpoint for nephrotoxicity induced by gardenia yellow based on an integrated strategy using bioinformatics analysis and in vitro testing validation.

To explore the potential the adverse outcome pathway of Gardenia Yellow (GY)-induced sensitive endpoint for nephrotoxicity, an integrated strategy was applied in the present study. Using bioinformatic analysis, based on the constructed Protein-protein interaction networks, Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis on the core target network were performed to illustrate the potential gene targets and signal pathways. Then, the most enriched pathway was validated with Cell counting kit-8 assays and Western blot analysis in embryonic kidney epithelial 293 cell models. According to the findings, GY may interact with 321 targets related to the endpoint. The five targets on the top ranking in the PPI network were STAT3, SRC, HRAS, AKT1, EP300. Among them, PI3K/Akt was the most enriched pathway. In vitro testing showed that GY exerted a proliferative effect on the cell variability in a dose-dependent manner. GY at concentration of 1000 µg/ml and stimulation for 30 min can significantly enhance the expression of phosphorylated Akt. Thus, after the quantitative weight of evidence evaluation, Akt phosphorylation induced PI3K/Akt activation was speculated as a molecular initiating event leading to a proliferative and inflammatory response in renal tubular epithelial cells.

[Experimental study of TGF-ß2 antisense oligonucleotide on inhibiting corneal scar hyperplasia in rabbits].

OBJECTIVE: To investigate the influence of TGF-ß2 antisense oligonucleotide (ASON) on preventing corneal scar hyperplasia in rabbits and to provide experimental evidence for its clinical application. METHODS: It was an experimental study. One hundred and ninety two New Zealand white rabbits were randomly divided into 4 groups (groups A, B, C and D). Corneal injury models were established in all groups. There were 48 experimental animals in each group. TGF-ß2 ASON was dropped into right eyes in group A, dexamethasone was dropped into right eyes in group B, deionized water was dropped into right eyes in group C and nothing was dropped into right eyes in group D after the operation. The corneas were surgically removed and assessed by hematoxylin eosin (HE) staining, immunohistochemical study and real time PCR at four different time points (4 d, 7 d, 14 d and 28 d) after surgery. RESULTS: HE staining: at the same time point, fibroblasts in the groups A and B were significantly fewer than that in the groups C and D, the difference was statistically significant (P < 0.05), but there was no significant difference between groups A and B or groups C and D. Immunohistochemical observation found that the expression of α-smooth muscle actin (α-SMA) positive fibroblasts could be observed by the 4th day (9.44 ± 0.47/HP), reached a climax by the 7th day (12.50 ± 0.81/HP), and returned to the baseline levels by the 14th day (0.85 ± 0.43/HP) in the group A, which was similar to that in the group B (9.49 ± 0.95, 12.42 ± 0.70, 0.86 ± 0.79/HP) at the same time point (P > 0.05), but it was significantly fewer than that in the group C(20.14 ± 0.78, 18.19 ± 1.28, 4.87 ± 0.58/HP) and group D(20.21 ± 0.92, 18.25 ± 1.39, 5.00 ± 2.217/HP), which was statistical significant (P < 0.05). The staining intensity of fibronectin (FN) in groups A and B was significantly weaker than that in groups C and D. Real time PCR analysis showed that at each time point, the expression of TGF-ß2 mRNAs in groups A and B was significantly lower than that in groups C and D (P < 0.05). CONCLUSIONS: TGF-ß2 ASON can effectively prevent the proliferation of corneal tissue by inhibiting the activity of TGF-ß2 after injury. The early stage of corneal repair is 7 days after injury, so it is important to use TGF-ß2 ASON at this stage to inhibit the scar hyperplasia. In addition, it is safe to apply TGF-ß2 ASON topically to protect the cornea from obvious side effects.

[Effect of invigorating lung and kidney prescription and its components on secretion of cytokines induced by CSE and LPS].

OBJECTIVE: To observe effect of Invigorating Lung and Kidney Prescription (ILKP) on secretion of cytokines induced by Lipopolysaccharide (LPS) and cigarette smoke extract (CSE) in vitro and discuss prescription regularity of ILKP and reveal molecular mechanism of ILKP treatment on COPD. METHODS: Alveolar epithelial cells A549, airway epithelial cells H292 and monocyte macrophage THP-1 were stimulated with LPS and CSE, in which samples were divided into normal group (20% normal rat serum), model group (20% normal rat serum and 5% CSE with 75 pg/mL LPS), ILKP group and its Components [20% Replenishing qi group (1), Replenishing Lung and Kidney group (2), Activating blood group (3), Reduce phlegm group (4), Replenishing qi with Replenishing Lung and Kidney group (5), Replenishing qi with Activating blood group (6), Replenishing qi with Reduce phlegm group (7), Replenishing qi with Replenishing Lung and Kidney and Activating blood group (8), Replenishing qi with Replenishing Lung and Kidney and Reduce phlegm group (9), Replenishing qi with Activating blood and Reduce phlegm group (10), ILKP serum group (11)]. The cytokines were detected by ELISA and the transcription factors of cytokines were analyzed with Network tool. RESULTS: Compared with the normal group,the secretion of MMP-9, GM-CSF and TGF-beta by THP-1 cells,TIMP by A549 cells,TNF-alpha, MMP-9, GM-CSF, CD36 and TIMP-1 by H292 cells all were increased, while secretion of TNF-alpha, PPAP2B, IL-3 and SOD decreased by THP-1 cells, MMP-9, TNF-alpha, TGF-beta and IL-3 by A549 cells, IL-8 by H292 cells all were decreased. Compared with model group, for THP-1 cells, the secretion of GM-CSF in 3, 6, 10 and 11 prescription groups,TGF-beta in 10 and 11 prescription groups all were decreased. MMP-9 decreased in 9 and 5 prescription groups while it increased in 1, 2, 3, 4, 7, 8, 10 and 11 prescription groups. TNF-alpha was increased in 1, 2, 3, 4, 6, 8, and 9 prescription groups, as it decreased in 11 prescription group. Except 2 prescription group, IL-3 was increased in all other drugs groups. SOD in 2, 4, 5, 8, 9, 11 prescription groups, and PPAP2B in 1, 2, 3, 4, 5, 6, 7 prescription groups were increased while PPAP2B was decreased in 11 prescription group. For A549 cells, the secretion of TIMP-1 was increased in 1, 2, 3, 4, and 5 prescription groups, while it was decreased in 11 prescription groups; Except 1 and 4 prescription group, MMP-9 was increased in other groups; Except TGF-beta was increased in 11 prescription groups, it was increased in other groups; IL-3 was increased in 1, 2, 4, 5, 11 and 3 prescription groups. For H292 cells, the secretion of GM-CSF in 5,6,7 and 2 prescription groups,TNF-alpha in 1, 2, 4, 6, 7, 8, 9 and 10 prescription groups, MMP-9 in 4, 8, 7 and 9 prescription groups all were decreased. CONCLUSION: The effect of ILKP and its components on secretion of cytokines induced by LPS and CSE and its target cells are different, in which Activating blood prescription focuses on inflammation cytokines, Replenishing Lung and Kidney prescription on Lipid metabolism and redox regulation. The effect of ILKPA is the most widely, followed by Activating blood Replenishing Lung and Kidney, and Reduce phlegm prescription on Protease regulation.

[Clinical observation of visual quality after aspherical intraocular lens implantation in traumatic cataract].

OBJECTIVE: To observe the visual quality of aspherical intraocular lens (IOL) implantation in traumatic cataract patients. METHODS: Prospective clinical study.96 traumatic cataract patients (96 eyes) suffered from penetrating corneal trauma chosen from the first affiliated hospital of Zhengzhou university during June 2009 to June 2012. They were divided into two groups based on the different type of intraocular lens. The experimental group (48 eyes) was implanted with aspherical IOL and the control group (48 eyes) was implanted with traditional sphere IOL.Uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), contrast sensitivity (CS) and stereoscopic vision were observed at 1 month, 3 months, and 6 months after surgery. At the same time, questionnaire survey about the satisfaction of patients was also performed. The t test was used to compare the preoperative general condition, postoperative visual acuity, contrast sensitivity and stereoscopic vision of the two groups, and the rank sum test was used to compare the astigmatism and the satisfaction of patients. RESULTS: There was no significant difference in UCVA (t = 1.37, 1.28,0.71, P > 0.05) between the experimental group (0.56 ± 0.22, 0.68 ± 0.13,0.84 ± 0.15) and the control group (0.51 ± 0.17, 0.61 ± 0.20,0.81 ± 0.17) at three time points. There was no significant difference in BCVA (t = 0.87, 1.38, 1.39, P > 0.05) between the experimental group (0.62 ± 0.13, 0.74 ± 0.21, 0.87 ± 0.10) and the control group (0.57 ± 0.25,0.69 ± 0.22,0.84 ± 0.15) . The same result happened in stereoscopic vision at 6 months after surgery (far stereopsis:123.5 ± 7.8 vs 126. 9 ± 5.9, t = 0.64, P > 0.05;near stereopsis:90.5 ± 7.8 vs 95.2 ± 3.5; t = 1.36, P > 0.05) between experimental group and control group. The contrast sensitivity of the experimental group in every stage (3 c/d:1.52 ± 0.18, 6 c/d:1.68 ± 0.19, 12 c/d:1.29 ± 0.14, 18 c/d:1.04 ± 0.20) was superior to the control group (3 c/d:1.49 ± 0.27, 6 c/d:1.57 ± 0.21, 12 c/d:1.14 ± 0.20, 18 c/d:0.85 ± 0.14) , especially on the glare sensitivity (the experimental group:3 c/d:1.40 ± 0.15, 6 c/d:1.52 ± 0.22, 12 c/d:1.21 ± 0.18, 18 c/d:0.91 ± 0.14, the control group:3 c/d:1.13 ± 0.13, 6 c/d:1.13 ± 0.28, 12 c/d:0.92 ± 0.13, 18 c/d:0.54 ± 0.16) Compared two groups of difference have statistical significance (free from glare:3 c/d:t = 2.829, 6 c/d:t = 4.092, 12 c/d:t = 3.055, 18 c/d:t = 2.093;glare:3 c/d:t = 2.650, 6 c/d:t = 3.105, 12 c/d:t = 3.395, 18 c/d:t = 2.215;P < 0.05) .Questionnaire survey showed the experimental group (72.9%) was statistically significantly higher (t = 3.016, P < 0.05) than that in the control group (54.1%) on the satisfaction of patients. CONCLUSIONS: The visual quality with implantation of aspherical IOL in traumatic cataract patients is superior to traditional sphere IOL. Aspherical IOL is more appropriate for the patients with small and peripheral corneal scar.It can reduce the visual function damage to minimum caused by trauma.

Amlodipine inhibits TNF-alpha production and attenuates cardiac dysfunction induced by lipopolysaccharide involving PI3K/Akt pathway.

Calcium channel blockers (CCBs) are widely used in the therapy of cardiovascular diseases. Recent studies have shown that several CCBs exerted distinct anti-inflammatory effect in myocardial dysfunction models. The purpose of the present study was to evaluate therapeutic effect and possible mechanism of action of amlodipine, one of the widely used CCBs, on rat cardiac dysfunction during sepsis induced by lipopolysaccharide (LPS). Pretreatment of the rats with amlodipine (10 or 30 mg/kg, i.v.) delayed the fall of mean arterial blood pressure caused by LPS. Amlodipine also significantly inhibited the elevation of plasma tumor necrosis factor alpha (TNF-alpha) and decreased levels of inducible nitric oxide synthase (iNOS) in response to LPS challenge. To investigate the mechanism of the action of amlodipine, neonatal rat cardiomyocytes were used as a model. Amlodipine concentration-dependently decreased the release of TNF-alpha and iNOS protein expression, and suppressed the degradation and phosphorylation of inhibitor of kappaB-alpha (IkappaB-alpha) in LPS-activated neonatal rat cardiomyocytes. Further studies revealed that amlodipine markedly activated phosphatidylinositiol 3-kinase (PI3K) and Akt, downstream of the PI3K signal cascade. Application of PI3K inhibitors, wortmannin and LY294002 attenuated the depression of TNF-alpha and iNOS expression by amlodipine in LPS-induced cardiomyocytes. These findings may explain some cardioprotective effects of amlodipine in LPS-mediated sepsis and suggest that the inhibition of TNF-alpha and iNOS expression by amlodipine is, at least in part, dependent on PI3K/Akt signaling pathway.

[Multiresidue analysis of organochlorine pesticides and the application to Chinese herbal medicine].

[A review of study on multiresidue analyses of organochlorine pesticides(OCPs) was presented here, which included sample extraction, clean-up and determination. The multiresidue analytical methods applied to Chinese herbal medicine was also discussed.

PC-407 inhibited proliferation and induced apoptosis in human colon cancer SW-1116 cells.

AIM: To study whether PC-407 [4-[5-naphthyl-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide] inhibits cell viability and induces apoptosis in human colon cancer SW-1116 cells. METHODS: Inhibition of SW-1116 proliferation was measured by MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope and electron microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. RESULTS: PC-407 inhibited SW-1116 cell proliferation in a concentration-dependent manner after 3 d of treatment, and the IC(50) for PC-407 inhibition of cell number was 16.67+/-0.17 micromol/L. After incubation of SW-1116 cells with PC-407 20 micromol/L for 24 h, morphological changes of typical apoptosis were observed by AO/EB staining or transmission electron microscopy. Flow cytometry analysis showed that PC-407 induced apoptosis in SW-1116 cells in a time- and concentration-dependent manner. The agarose gel electrophoresis of DNA revealed a ladder pattern 48 h later. CONCLUSION: PC-407 inhibited proliferation and induced apoptosis in the human colon cancer SW-1116 cell line.