Objective: Using network pharmacology and molecular docking technology along with scutellarein (SE) as a reference, this study predicted the anti-cerebral ischemia targets of both baicalein (BE) and genipin (GE). It is hoped that these will provide a reference for clinical prevention and development of ischemic diseases. Methods: SE, BE and GE targets were predicted using TCMSP, Swiss Target Prediction, Stitch database search and literature mining methods. Targets related to cerebral ischemia diseases could be predicted by DisGeNET, CTD, NCBI Gene, OMIM, DrugBank and PharmGkb databases. Cytoscape 3.3.0 was used to construct the small molecule-target network. GO function enrichment and KEGG pathway analysis of SE, BE and GE specific anti-cerebral ischemia targets were analyzed with the DAVID database. Autodock Vina software was used for molecular docking, testing the binding energy of BE, GE and SE to targets of cerebral ischemia. The optimal target protein was selected according to the binding energy and inhibition concentration of receptor and ligand. Results: A total of 30 potential targets of SE, 59 potential targets of BE and 35 potential targets of GE were found. Common anti-cerebral ischemia targets of SE and BE were PIK3CG, CYP1A2, VEGFA, ALOX5 and PTGS2, while common anti-cerebral ischemia targets of SE and GE were PTGS2. Molecular docking results demonstrated that the binding energy and inhibitory concentration of receptor PTGS2 to the three drugs were relatively low. Enrichment of GO function showed that common targets of BE-SE were mainly distributed in cytoplasm, organelle membrane, endoplasmic reticulum and other elements. These elements had binding functions with metal ions and cations, catalytic and oxidoreductase activities, and they participated in cell lipid, carboxylic acid, oxygenic acid, organic acid metabolism and fatty acid synthesis. Results: of the KEGG analysis demonstrated that receptor PTGS2 mainly acted on the arachidonic acid metabolism pathway and the vascular endothelial growth factor signaling pathway. A combination of BE and GE functioned in the treatment of cerebral ischemia disease by inhibiting expression of PTGS2 (COX-2) and vascular endothelial growth factor protein, thus reducing brain injury caused by inflammatory factors and improving the permeability of the blood brain barrier (BBB). Conclusion: This study predicted potential targets of BE and GE compatibility in the treatment of cerebral ischemia diseases and preliminarily verified mechanisms of action in the treatment of cerebral ischemia diseases. It provides valuable data for further study into the mechanisms of BE and GE compatibility for the treatment of cerebral ischemia as well as developing a basis for the next synthesis of new derivatives.

Objective To explore clinical features and drug sensitivity of Streptococcus pneumoniae (SP) isolated from pediatric patients with lower respiratory tract infection, and to provide evidence for clinical use of antibiotics. Methods A total of 6 358 clinical SP isolates from children with lower respiratory tract infection from January 2008 to December 2012 were col-lected and retrospectively analyzed. The antibiotic sensitivity was done by Kirby-Bauer method and E-test, and all results were in strict accordance with the rules of CLSI. Results The isolated SP strains were mainly from different departments of pediatrics. All clinical cases with SP infection mainly included pneumonia and bronchitis. The resistance rates of 6 358 SP strains to penicil-lin, cefotaxime, erythromycin, clindamycin, cotrimoxazole, tetracycline, chloramphenicol, levolfoxacin, vancomycin were 5.0%, 12.9%, 98.7%, 96.0%, 92.2%, 7.3%, 5.6%, 0.2%and 0.0%respectively, and the resistance rate to penicillin and cefotaxime was signiifcantly different in every years (all P<0.05). The resistance rates of the 318 penicillin-resistant SP strains to the above anti-biotics were 100.0%, 42.6%, 100.0%, 100.0%, 99.2%, 23.6%, 6.8%, 0.6%, 0.0%respectively, and the resistance rate to penicillin and cefotaxime was signiifcantly different (P=0.001). Conclusions The antibiotic resistance rates of SP strains isolated from children with lower respiratory tract infection were higher to erythromycin, clindamycin, cotrimoxazole and tetracycline, and an increasing rate in drug resistance to cefotaxime was observed in recent years. Appropriate antibiotics should be selected for the treatment of infection according to drug sensitivity.

Objective To explore the regulation mechanism of autophagy-related protein, microtubule-associated protein 1 light chain 3 (LC3), via c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cell lines. Methods Human choriocarcinoma JEG-3 cell lines, and methotrexate resistant choriocarcinoma JEG-3 (JEG-3/MTXR) cell lines were used in our present study. Phosphorylation c-Jun (p-c-Jun) was evaluated after exposure to 0.02 ng/ml methotrexate for 72 hours in both cells by western blot. c-Jun gene was knockdown by small interference RNA (siRNA) in JEG-3/MTXR cells, and LC3 was evaluated by western blot and reverse transcription-PCR. The binding of LC3 promoter with c-Jun protein was detected via chromatin immunoprecipitation assay (ChIP) with or without 0.02 ng/ml methotrexate exposure. Results The results showed that p-c-Jun was up-regulated after methotrexate treatment for 72 hours (1.99±0.20, versus 0.20±0.06 at 0 hour;P<0.05) by western blot analysis in JEG-3/MTXR cell lines. Further investigation demonstrated that c-Jun-siRNA could inhibit the up-regulation of LC3 formation and after methotrexate exposure (LC3 mRNA:1.24±0.17 versus 3.03±0.43;LC3 protein:0.52±0.07 verus 1.20± 0.15; all P<0.05). The binding of LC3 promoter by c-Jun protein was up-regulated after methotrexate treatment by the method of ChIP in methotrexate resistant JEG-3/MTXR cells [(2.95 ± 0.35) times]. Conclusion Autophagy-related gene LC3 expression regulated by c-Jun protein may be involved in the effect mechanism of the development of methotrexate resistance in choriocarcinoma JEG-3 cells.

Objective To investigate the effect of combined Metformin and esomeprazole therapy on plasma levels of gastrin,blood glucose,glycosylated hemoglobin(HbA1c) and insulin in elderly patients with type 2 diabetes.Methods A randomized,double-blind,placebo-controlled study of 41 elderly patients with type 2 diabetes was conducted.Patients were randomly assigned into treatment group(combination therapy with Metformin 0.5 g,bid or tid and Esomeprazole 20 mg,qd,for 12 weeks)and placebo group(Metformin monotherapy 0.5 g,bid or tid,for 12 weeks).Fasting blood samples were taken from vein before and after treatment.Fasting serum levels of gastrin,glucose,HbA1c,insulin,lipids,liver and renal functions were compared between the two groups.The homeostasis model of assessment for insulin resistance index(HOMA-IR) and insulin secretion index (HOMA-β)were calculated,and complications were recorded.Results There were no significant differences in body mass index and waist circumference between the two groups.Serum gastrin level was slightly increased in the treatment group 12 weeks after treatment,but without statistically significance [(127.20±9.21)ng/L vs.(131.53±7.84)ng/L,P＞0.05],and serum gastrin level had no significant differences in the placebo group before and after treatment [(128.42±4.58)ng/L vs.(127.51±3.47)ng/L,P＞0.05].However,there were no significant differences in the changes of blood glucose,HbA1c,insulin,HOMA-β and HOMA-IR before versus after therapy,and between the two group(all P＞0.05).Conclusions Combined Metformin and insulin therapy cannot increase serum gastrin and insulin levels and has no significant effect on reducing blood glucose and HbA1c levels in elderly patients with type 2 diabetes.

Abdominal Injuries ; complications ; Accidents, Traffic ; Humans ; Male ; Mediastinum ; Middle Aged ; Splenosis ; etiology ; pathology ; surgery ; Thoracotomy

Background Posterior capsular opacification(PCO) is common complication after extrecapsular extract of cataract.Matrix metalloproteinases-3 (MMP-3) can degrade all the extracellular matrix except polyose.The gene therapy of PCO upon MMP-3 is the researching hot topic.Fibronectin ( FN ) is a degrade gelatin,so its expression can reflect the effect of MMP-3 on LECs indirectly. Objective The aim of this study was to construct MMP-3 eukaryotic recombination plasmid and transfect to lens epithelium cells(LECs) for the observation of MMP3 expression,and to explore the feasibility of gene therapy for after cataract. Methods Six fresh lenses were obtained from pigs.LECs were cultured using explant method.The eukaryotic expression vector pEGFP-N1-MMP-3 was reconstructed with MMP-3 and pEGFP-N1 plasmids.The accuracy of MMP-3 gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-MMP-3 into LECs of pig,the expression of MMP-3 protein in the cells was indirectly observed by green fluorescent protein.The expression of FN in LECs was detected using Western blot. Results The result of double enzyme digestion was consistent with the base number of pEGFP-N1 plasmids and target fragment.By enlacing the result of DNA sequencing analysis with software,the resemblance of the DNA sequence of MMP-3 from recombination plasmid pEGFP-N1-MMP-3 and that of homo MMP-3 was 99.6％,indicating that the target fragment was inserted to pEGFP-N1 plasmids successfully.Green fluorescence for GFP was seen in the LECs in pEGFP-N1-MMP-3 transfected group,but absent response for GFP was in empty vector group.Western blot revealed that the relative expression level of FN in LECs was 0.666±0.008 in pEGFP-N1-MMP-3 trasfected group and 0.326 ±0.071 in empty vector group,with a significant difference between these two groups(P=0.000). Conclusions Eukaryotic recombination plasmid pEGFP-N1-MMP-3 is successfully constructed,and MMP-3 can be expressed in LECs after transfected.These results lay a foundation for the further research of MMP-3 gene therapy for PCO.

Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.

LC-Q-TOF-MS and LC-IT-MS in positive and negative ion mode were applied to simultaneously characterize the constituents in Suanzaoren tang. Analysis was performed on an Agilent Zorbax SB-C18, Rapid Resolution HT column(4.6 mmx 50 mm, 1. 8 micro m) with gradient elution of acetonitrile(A) -aqueous solution containing 0. 05% formic acid(B) at a flow rate of 0. 6 mL min(-1) and the column temperature was 30 degreesC. By comparing MS fragmentation, accurate molecular weight, literature date and standard compounds information, a total of48 compounds were successfully identified or speculated. The origins of these compounds were assigned to the corresponding Chinese medicine. Thirty-one compounds were reported in Suanzaoren tang for the first time. LC-Q-TOF-MS combined with LC-IT-MS is a simple and rapid tool for the identification of constituents of Suanzaoren tang, and the results could provide evidence for the research on quality combined and effective constituents of Suanzaoren tang.
Chromatography, Liquid ; Drugs, Chinese Herbal ; chemistry ; Mass Spectrometry ; Organic Chemicals ; analysis

Objective To explore the feasibility,advantages and disadvantages of combined trans- gastric and transcolonic routes for NOTES.Methods A female swine was used in this study.Transgastric entrance was the first,followed by transeolonie entrance.A dual-channel endoscope was inserted through the porcine mouth into the gastric cavity and penetrated into the peritoneal cavity through the puncture and bal- loon dilatation of the gastric wall.Then under direct visualization through the transgastric approach,the other endoseope was advanced into the peritoneum.Using the two endoscopes inside the peritoneal cavity,collabo- rative peritoneoseopy was performed by the two endoscopists.After the examination the incisions in the stom- ach and the colon wall were closed with Endoclips.The animal was sacrificed for post-mortem examination with particular attention to the entrance sites and presence of any complications related to the access or to ma- nipulations inside the peritoneal cavity.Results No hemorrhage oecurred during the puneture and balloon dilatation or bow-knife cutting of the gastric wall or the eolonic wall.The liver was damaged while a needle knife penetrated the gastric wall.On the contrary,no organs were damaged during the needle knife penetra- ted the eolonic wall under direet visualization through the transgastric approach.It was difficult to find the gallbladder or the oviduct with a"single arm",but it was easy to see them with the double routes.It was easier to close the colonic incision than to close the gastrie wall with Endoclips.Conclusion Combined transgastric and transcolonic route for NOTES is feasible and it seems to be easier to show a target compared with a single route.

Objective·To establish a cell-based screening system for identification of compounds with activity in regulating retinoic acid receptor (RARα) stability. Methods·The modified pMSCV plasmid constructs, named as RARα-EGFP-IRES-DsRed, consists of enhanced green fluorescent protein (EGFP) fusing to RARα and red fluorescent protein (DsRed) as internal references incorporating the internal ribosome entry site (IRES) as interval sequence. The RARα-EGFP-IRES-DsRed plasmid was stably transfected into NB4 cells which were named as NB4-pMGIR-RARα. Fluorescence signals of EGFP and DsRed indirectly reflecting the expression of RARα, were detected by flow cytometry in cells that were treated with all-trans retinoic acid, sodium valproate, cytarabine, lenalidomide, etoposide, montelukast and gambogic acid, respectively. Effects of these compounds on the expression of RARα protein were further examined by Western blotting. Results·A double fluorescence reporter system for screening compounds that can increase the stability of RARα protein was successfully established, and sodium valproate was identified as a potent compound to promote the stability of RARα. Conclusion·The double fluorescence reporter system can be used to screen compounds regulating the stability of RARα protein, which can be further used to identify compounds regulating the stability of other proteins.